U.S. patent application number 13/919916 was filed with the patent office on 2014-01-02 for linear polyester and semi-linear glycidol polymer systems: formulation and synthesis of novel monomers and macromolecular structures.
This patent application is currently assigned to Vanderbilt University. The applicant listed for this patent is Vanderbilt University. Invention is credited to Dain B. Beezer, Eva M. Harth, GuangZhao Li, Benjamin R. Spears, David M. Stevens.
Application Number | 20140005278 13/919916 |
Document ID | / |
Family ID | 49778767 |
Filed Date | 2014-01-02 |
United States Patent
Application |
20140005278 |
Kind Code |
A1 |
Harth; Eva M. ; et
al. |
January 2, 2014 |
LINEAR POLYESTER AND SEMI-LINEAR GLYCIDOL POLYMER SYSTEMS:
FORMULATION AND SYNTHESIS OF NOVEL MONOMERS AND MACROMOLECULAR
STRUCTURES
Abstract
Disclosed herein are glycidol-based polymers, nanoparticles, and
methods related thereto useful for drug delivery. This abstract is
intended as a scanning tool for purposes of searching in the
particular art and is not intended to be limiting of the present
invention.
Inventors: |
Harth; Eva M.; (Nashville,
TN) ; Beezer; Dain B.; (Nashville, TN) ; Li;
GuangZhao; (Nashville, TN) ; Spears; Benjamin R.;
(Nashville, TN) ; Stevens; David M.; (Nashville,
TN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Vanderbilt University |
Nashville |
TN |
US |
|
|
Assignee: |
Vanderbilt University
Nashville
TN
|
Family ID: |
49778767 |
Appl. No.: |
13/919916 |
Filed: |
June 17, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
61660675 |
Jun 15, 2012 |
|
|
|
Current U.S.
Class: |
514/772.7 ;
525/408; 525/451; 526/273; 528/220; 528/354; 528/361; 528/393;
528/409; 528/421 |
Current CPC
Class: |
A61K 47/34 20130101;
A61K 9/5031 20130101; C08G 65/3348 20130101; C08G 65/00 20130101;
C08G 65/3326 20130101; A61K 9/0019 20130101; C08G 65/22 20130101;
C08G 65/34 20130101; C08G 64/42 20130101; A61K 47/32 20130101; C08G
65/2615 20130101; C08G 65/002 20130101 |
Class at
Publication: |
514/772.7 ;
528/421; 528/393; 526/273; 528/361; 528/354; 528/220; 528/409;
525/451; 525/408 |
International
Class: |
A61K 47/34 20060101
A61K047/34; A61K 47/32 20060101 A61K047/32 |
Claims
1. A polymer comprising repeating units selected from: ##STR00183##
wherein R.sup.0 is selected from H, alkyl, NH.sub.2, and R.sup.1;
wherein R.sup.1 comprises a crosslinking functionality; wherein
repeating units A1, A2, B1, and B2 account for at least about 50
wgt % of the polymer; and wherein the ratio of (A1+A2):(B1+B2) is
greater than 1.
2. The polymer of claim 1, further comprising at least one
repeating unit formed from a monomer selected from: ##STR00184## or
a combination thereof.
3. The polymer of claim 2, wherein the polymer comprises at least
one repeating unit from a monomer selected from: ##STR00185## or a
combination thereof, and wherein the polymer is oxidized to form
repeating units comprising epoxides or alkynes.
4. The polymer of claim 1, further comprising crosslinks, wherein
the crosslinks comprises ##STR00186## wherein at least one of
##STR00187## is not 0.
5. The polymer of claim 4, wherein the crosslinks comprises one or
more of ##STR00188## or any combination thereof.
6. The polymer of claim 4, wherein the polymer comprises
##STR00189## ##STR00190## ##STR00191## ##STR00192##
##STR00193##
7. A nanoparticle comprising the polymer of claim 1.
8. The nanoparticles of claim 7, wherein the polymer is crosslinked
via crosslinks, wherein the crosslinks comprises ##STR00194##
wherein at least one of ##STR00195## is not 0.
9. A method for making a polymer, the method comprising the step of
polymerizing glycidol in the presence of a tin catalyst.
10. The method of claim 9, wherein the tin catalyst is
Sn(OTf).sub.2.
11. The method of claim 9, wherein the resultant polymer comprises
repeating units selected from: ##STR00196## wherein R.sup.0 is
selected from H, alkyl, NH.sub.2, and R.sup.1; wherein R.sup.1
comprises a crosslinking functionality; wherein repeating units A1,
A2, B1, and B2 account for at least about 50 wgt % of the polymer;
and wherein the ratio of (A1+A2):(B1+B2) is greater than 1.
12. The method of claim 9, wherein the resultant polymer further
comprises at least one repeating unit formed from a monomer
selected from: ##STR00197## or a combination thereof.
13. The method of claim 9, further comprising the step of
crosslinking the polymer with crosslinks, wherein the wherein the
crosslinks comprises ##STR00198## wherein at least one of
##STR00199## is not 0.
14. The method of claim 9, wherein the polymerization step is
performed at a temperature of from -80.degree. C. to 50.degree.
C.
16. A method of forming a nanoparticle comprising: (a) providing a
polymer of claim 3; and (b) crosslinking the polymer with
crosslinks, wherein the crosslinks comprises ##STR00200## wherein
at least one of ##STR00201## is not 0, thereby forming a
nanoparticle.
17. A drug delivery method comprising the step of administering to
a subject a composition comprising the polymer of claim 1, in
combination with at least one pharmaceutically active agent and/or
biologically active agent.
18. A pharmaceutical composition comprising (a) the compound of
claim 1; (b) pharmaceutically active agent and/or biologically
active agent; and (c) a pharmaceutically acceptable carrier.
19. A composition comprising (a) a polymer comprising repeating
units formed from monomers ##STR00202## wherein the polymer is
crosslinked via crosslinks, wherein the crosslinks comprises
##STR00203## wherein at least one of ##STR00204## is not 0; and (b)
a polymer of claim 1, wherein the polymer of claim 1 is optionally
covalently bonded to the polymer.
20. A polymer comprising a structure formed from reacting a polymer
of claim 1 further comprising repeating units ##STR00205## with a
polymer comprising at least one repeating unit formed from
##STR00206##
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional
Application No. 61/660,675 filed on Jun. 15, 2012; which is
incorporated herein by reference in its entirety.
BACKGROUND
[0002] Many highly efficacious drugs have already been created and
the main hurdle that these drug molecules have to overcome is their
hydrophobicity. Due to this lack of solubility, regardless of the
drugs efficacy, the molecules will never be cleared as viable
treatment options. Furthermore, biological therapeutics such as
antibodies and proteins (e.g., growth factors) are not stable for a
prolonged time in the biological environment and impedes their
activity and therapeutic efficacy. Moreover, it has been found that
therapeutics can work very efficiently together and enhance the
therapeutic outcome known as the synergistic effect.
[0003] Thus, there remains a need for delivery systems that address
hydrophobicity and/or lack of solubility. In view of the need of
delivery systems that deliver drugs of different nature, can
control the kinetics of the delivery, and react to external
stimuli, multifaceted delivery systems are being developed. The
combinations of 3-D nanoparticles are designed to deliver small
molecules that are imbedded in non-crosslinked or crosslinked
matrices are of interest. Additionally, 2-D materials that contain
no 3-D nanoparticle materials are crosslinked to hydrophilic
networks to be formed in click reactions in hydrophilic and
hydrophobic environments. The functionalities in these hydrogel
materials allow response to heat, reconfiguring the network but not
destroying the structure.
SUMMARY OF THE INVENTION
[0004] In accordance with the purpose(s) of the invention, as
embodied and broadly described herein, the invention, in one
aspect, relates to compounds that can be used in drug delivery, and
composition thereof and methods thereof.
[0005] Disclosed herein is a polymer comprising repeating units
selected from:
##STR00001##
wherein R.sup.0 is selected from H, alkyl, NH.sub.2, and R.sup.1;
wherein R.sup.1 comprises a crosslinking functionality; wherein
repeating units A1, A2, B1, and B2 account for at least about 50
wgt % of the polymer; and wherein the ratio of (A1+A2):(B1+B2) is
greater than 1.
[0006] Also disclosed herein is a nanoparticle comprising the
disclosed compounds.
[0007] Also disclosed is a method for making a polymer, the method
comprising the step of polymerizing glycidol in the presence of a
tin catalyst.
[0008] Also disclosed herein is a method for forming a nanoparticle
comprising: a. providing a polymer disclosed herein and
crosslinking polymer with crosslinks disclosed herein.
[0009] Also disclosed is a drug delivery method comprising the step
of administering to a subject a composition comprising a polymer or
nanoparticle disclosed herein, in combination with at least one
pharmaceutically active agent and/or biologically active agent.
[0010] Also disclosed herein is a pharmaceutical composition
comprising a polymer or nanoparticle disclosed herein; a
pharmaceutically active agent and/or biologically active agent; and
a pharmaceutically acceptable carrier.
[0011] While aspects of the present invention can be described and
claimed in a particular statutory class, such as the system
statutory class, this is for convenience only and one of skill in
the art will understand that each aspect of the present invention
can be described and claimed in any statutory class. Unless
otherwise expressly stated, it is in no way intended that any
method or aspect set forth herein be construed as requiring that
its steps be performed in a specific order. Accordingly, where a
method claim does not specifically state in the claims or
descriptions that the steps are to be limited to a specific order,
it is no way intended that an order be inferred, in any respect.
This holds for any possible non-express basis for interpretation,
including matters of logic with respect to arrangement of steps or
operational flow, plain meaning derived from grammatical
organization or punctuation, or the number or type of aspects
described in the specification.
BRIEF DESCRIPTION OF THE FIGURES
[0012] The accompanying Figures, which are incorporated in and
constitute a part of this specification, illustrate several aspects
and together with the description serve to explain the principles
of the invention.
[0013] FIG. 1 shows schematic representation of drug-loaded
hyperbranched polyglycerols.
[0014] FIG. 2 shows a fluorescent image of polyglycerols loaded
with a therapeutic cargo.
[0015] FIG. 3 shows a schematic representation of degradability of
glycidol polymers.
[0016] FIG. 4 shows schematic representations of glycidol polymers
carrying solubilized biological cargo.
[0017] FIG. 5 shows a schematic representation of a poly(ethylene
glycol)-protein conjugate.
[0018] FIG. 6 shows a schematic representation of a reaction scheme
for the introduction of allyl functionalities in the polymer
systems of the present invention.
[0019] FIG. 7 shows the equation for degree of branching in the
resultant polymers with the variables referring to the integration
values obtained from quantitative .sup.13CNMR investigation of the
present invention.
[0020] FIG. 8 shows .sup.13C-NMR spectra of glycidol homopolymer of
the present invention.
[0021] FIG. 9 shows a table reporting experimental NMR data and
degree of branching for polyglycidol systems of the present
invention.
[0022] FIG. 10 shows a visual representation of depression of
dendritic peak through kinetically controlled reactions of the
present invention.
[0023] FIG. 11 shows a visual representation of poly(glycidol)
branching possibilities.
[0024] FIG. 12 shows NMR spectra and a visual representation of
poly(glycidol) branching possibilities.
[0025] FIG. 13 shows a schematic representation of ring opening
possibilities for polyglycidol systems.
[0026] FIG. 14 shows NMR spectra for glycidyl ester allyl of the
present invention.
[0027] FIG. 15 shows NMR spectra for GLY/GEA polymer.
[0028] FIG. 16 shows a schematic representation of hydrophobicity
of glycidol polymers due to the presence of OPD.
[0029] FIG. 17 shows a schematic representation of a siRNA
complexation reaction.
[0030] FIG. 18 shows a schematic representation of a dendritic
polyglycerols with an amine shell.
[0031] FIG. 19 shows a schematic representation of nanoparticle
formation through a controlled thiolene-click reaction of the
present invention.
[0032] FIG. 20 shows a schematic representation of formation of
polyester nanoparticles.
[0033] FIG. 21 shows a schematic representation of thiolene-click
GLY/AGE nanoparticle formation of the present invention.
[0034] FIG. 22 shows a schematic representation of loading of
nanoparticle with small molecule drugs.
[0035] FIG. 23 shows a visual representation of the minimal size
dispersity of nanoparticle structures of the present invention.
[0036] FIG. 24 shows a transmission electron microscopy (TEM) image
for GLY/AGE nanoparticles of the present invention.
[0037] FIG. 25 shows a transmission electron microscopy (TEM) image
for GLY/AGE nanoparticles of the present invention.
[0038] FIG. 26 shows a transmission electron microscopy (TEM) image
for GLY/AGE nanoparticles of the present invention.
[0039] FIG. 27 shows a schematic representation of an exemplary two
component delivery system of the present invention.
[0040] FIG. 28 shows a schematic representation of exemplary
reconfigurable and responsive network systems of the present
invention.
[0041] FIG. 29 shows a schematic representation of exemplary
reaction schemes for formation of network systems of the present
invention.
[0042] FIG. 30 shows a visual representation of NMR spectra of an
exemplary glycidol polymer of the present invention.
[0043] Additional advantages of the invention will be set forth in
part in the description which follows, and in part will be obvious
from the description, or can be learned by practice of the
invention. The advantages of the invention will be realized and
attained by means of the elements and combinations particularly
pointed out in the appended claims. It is to be understood that
both the foregoing general description and the following detailed
description are exemplary and explanatory only and are not
restrictive of the invention, as claimed.
DESCRIPTION
[0044] The present invention can be understood more readily by
reference to the following detailed description of the invention
and the Examples included therein.
[0045] Before the present compounds, compositions, articles,
systems, devices, and/or methods are disclosed and described, it is
to be understood that they are not limited to specific synthetic
methods unless otherwise specified, or to particular reagents
unless otherwise specified, as such may, of course, vary. It is
also to be understood that the terminology used herein is for the
purpose of describing particular aspects only and is not intended
to be limiting. Although any methods and materials similar or
equivalent to those described herein can be used in the practice or
testing of the present invention, example methods and materials are
now described.
[0046] All publications mentioned herein are incorporated herein by
reference to disclose and describe the methods and/or materials in
connection with which the publications are cited. The publications
discussed herein are provided solely for their disclosure prior to
the filing date of the present application. Nothing herein is to be
construed as an admission that the present invention is not
entitled to antedate such publication by virtue of prior invention.
Further, the dates of publication provided herein can be different
from the actual publication dates, which can require independent
confirmation.
A. DEFINITIONS
[0047] As used herein, nomenclature for compounds, including
organic compounds, can be given using common names, IUPAC, IUBMB,
or CAS recommendations for nomenclature. When one or more
stereochemical features are present, Cahn-Ingold-Prelog rules for
stereochemistry can be employed to designate stereochemical
priority, E/Z specification, and the like. One of skill in the art
can readily ascertain the structure of a compound If given a name,
either by systemic reduction of the compound structure using naming
conventions, or by commercially available software, such as
CHEMDRAW.TM. (Cambridgesoft Corporation, U.S.A.).
[0048] As used in the specification and the appended claims, the
singular forms "a," "an" and "the" include plural referents unless
the context clearly dictates otherwise. Thus, for example,
reference to "a functional group," "an alkyl," or "a residue"
includes mixtures of two or more such functional groups, alkyls, or
residues, and the like.
[0049] Ranges can be expressed herein as from "about" one
particular value, and/or to "about" another particular value. When
such a range is expressed, a further aspect includes from the one
particular value and/or to the other particular value. Similarly,
when values are expressed as approximations, by use of the
antecedent "about," it will be understood that the particular value
forms a further aspect. It will be further understood that the
endpoints of each of the ranges are significant both in relation to
the other endpoint, and independently of the other endpoint. It is
also understood that there are a number of values disclosed herein,
and that each value is also herein disclosed as "about" that
particular value in addition to the value itself. For example, if
the value "10" is disclosed, then "about 10" is also disclosed. It
is also understood that each unit between two particular units are
also disclosed. For example, if 10 and 15 are disclosed, then 11,
12, 13, and 14 are also disclosed.
[0050] References in the specification and concluding claims to
parts by weight of a particular element or component in a
composition denotes the weight relationship between the element or
component and any other elements or components in the composition
or article for which a part by weight is expressed. Thus, in a
compound containing 2 parts by weight of component X and 5 parts by
weight component Y, X and Y are present at a weight ratio of 2:5,
and are present in such ratio regardless of whether additional
components are contained in the compound.
[0051] A weight percent (wt. %) of a component, unless specifically
stated to the contrary, is based on the total weight of the
formulation or composition in which the component is included.
[0052] As used herein, the terms "optional" or "optionally" means
that the subsequently described event or circumstance can or cannot
occur, and that the description includes instances where said event
or circumstance occurs and instances where it does not.
[0053] As used herein, the term "analog" refers to a compound
having a structure derived from the structure of a parent compound
(e.g., a compound disclosed herein) and whose structure is
sufficiently similar to those disclosed herein and based upon that
similarity, would be expected by one skilled in the art to exhibit
the same or similar activities and utilities as the claimed
compounds, or to induce, as a precursor, the same or similar
activities and utilities as the claimed compounds.
[0054] As used herein, the term "subject" refers to the target of
administration, e.g., an animal, such as a human. Thus the subject
of the herein disclosed methods can be a vertebrate, such as a
mammal, a fish, a bird, a reptile, or an amphibian. Alternatively,
the subject of the herein disclosed methods can be a human,
non-human primate, horse, pig, rabbit, dog, sheep, goat, cow, cat,
guinea pig or rodent. The term does not denote a particular age or
sex. Thus, adult and newborn subjects, as well as fetuses, whether
male or female, are intended to be covered. In one aspect, the
subject is a mammal. A patient refers to a subject afflicted with a
disease or disorder. The term "patient" includes human and
veterinary subjects. In some aspects of the disclosed methods, the
subject has been diagnosed with a need for treatment of one or more
muscle disorders prior to the administering step. In some aspects
of the disclosed method, the subject has been diagnosed with a need
for increasing muscle mass prior to the administering step. In some
aspects of the disclosed method, the subject has been diagnosed
with a need for increasing muscle mass prior to the administering
step.
[0055] As used herein, the phrase "identified to be in need of
treatment for a disorder," or the like, refers to selection of a
subject based upon need for treatment of the disorder. For example,
a subject can be identified as having a need for treatment of a
disorder (e.g., a disorder related to cancer) based upon an earlier
diagnosis by a person of skill and thereafter subjected to
treatment for the disorder. It is contemplated that the
identification can, in one aspect, be performed by a person
different from the person making the diagnosis. It is also
contemplated, in a further aspect, that the administration can be
performed by one who subsequently performed the administration.
[0056] As used herein, the terms "administering" and
"administration" refer to any method of providing a pharmaceutical
preparation to a subject. Such methods are well known to those
skilled in the art and include, but are not limited to, oral
administration, transdermal administration, administration by
inhalation, nasal administration, topical administration,
intravaginal administration, ophthalmic administration, intraaural
administration, intracerebral administration, rectal
administration, sublingual administration, buccal administration,
and parenteral administration, including injectable such as
intravenous administration, intra-arterial administration,
intramuscular administration, and subcutaneous administration.
Administration can be continuous or intermittent. In various
aspects, a preparation can be administered therapeutically; that
is, administered to treat an existing disease or condition. In
further various aspects, a preparation can be administered
prophylactically; that is, administered for prevention of a disease
or condition.
[0057] As used herein, the terms "effective amount" and "amount
effective" refer to an amount that is sufficient to achieve the
desired result or to have an effect on an undesired condition. For
example, a "therapeutically effective amount" refers to an amount
that is sufficient to achieve the desired therapeutic result or to
have an effect on undesired symptoms, but is generally insufficient
to cause adverse side effects. The specific therapeutically
effective dose level for any particular patient will depend upon a
variety of factors including the disorder being treated and the
severity of the disorder; the specific composition employed; the
age, body weight, general health, sex and diet of the patient; the
time of administration; the route of administration; the rate of
excretion of the specific compound employed; the duration of the
treatment; drugs used in combination or coincidental with the
specific compound employed and like factors well known in the
medical arts. For example, it is well within the skill of the art
to start doses of a compound at levels lower than those required to
achieve the desired therapeutic effect and to gradually increase
the dosage until the desired effect is achieved. If desired, the
effective daily dose can be divided into multiple doses for
purposes of administration. Consequently, single dose compositions
can contain such amounts or submultiples thereof to make up the
daily dose. The dosage can be adjusted by the individual physician
in the event of any contraindications. Dosage can vary, and can be
administered in one or more dose administrations daily, for one or
several days. Guidance can be found in the literature for
appropriate dosages for given classes of pharmaceutical products.
In further various aspects, a preparation can be administered in a
"prophylactically effective amount"; that is, an amount effective
for prevention of a disease or condition.
[0058] The term "pharmaceutically acceptable" describes a material
that is not biologically or otherwise undesirable, i.e., without
causing an unacceptable level of undesirable biological effects or
interacting in a deleterious manner.
[0059] As used herein, the term "derivative" refers to a compound
having a structure derived from the structure of a parent compound
(e.g., a compound disclosed herein) and whose structure is
sufficiently similar to those disclosed herein and based upon that
similarity, would be expected by one skilled in the art to exhibit
the same or similar activities and utilities as the claimed
compounds, or to induce, as a precursor, the same or similar
activities and utilities as the claimed compounds. Exemplary
derivatives include salts, esters, amides, salts of esters or
amides, and N-oxides of a parent compound.
[0060] As used herein, the term "pharmaceutically acceptable
carrier" refers to sterile aqueous or nonaqueous solutions,
dispersions, suspensions or emulsions, as well as sterile powders
for reconstitution into sterile injectable solutions or dispersions
just prior to use. Examples of suitable aqueous and nonaqueous
carriers, diluents, solvents or vehicles include water, ethanol,
polyols (such as glycerol, propylene glycol, polyethylene glycol
and the like), carboxymethylcellulose and suitable mixtures
thereof, vegetable oils (such as olive oil) and injectable organic
esters such as ethyl oleate. Proper fluidity can be maintained, for
example, by the use of coating materials such as lecithin, by the
maintenance of the required particle size in the case of
dispersions and by the use of surfactants. These compositions can
also contain adjuvants such as preservatives, wetting agents,
emulsifying agents and dispersing agents. Prevention of the action
of microorganisms can be ensured by the inclusion of various
antibacterial and antifungal agents such as paraben, chlorobutanol,
phenol, sorbic acid and the like. It can also be desirable to
include isotonic agents such as sugars, sodium chloride and the
like. Prolonged absorption of the injectable pharmaceutical form
can be brought about by the inclusion of agents, such as aluminum
monostearate and gelatin, which delay absorption. Injectable depot
forms are made by forming microencapsule matrices of the drug in
biodegradable polymers such as polylactide-polyglycolide,
poly(orthoesters) and poly(anhydrides). Depending upon the ratio of
drug to polymer and the nature of the particular polymer employed,
the rate of drug release can be controlled. Depot injectable
formulations are also prepared by entrapping the drug in liposomes
or microemulsions which are compatible with body tissues. The
injectable formulations can be sterilized, for example, by
filtration through a bacterial-retaining filter or by incorporating
sterilizing agents in the form of sterile solid compositions which
can be dissolved or dispersed in sterile water or other sterile
injectable media just prior to use. Suitable inert carriers can
include sugars such as lactose. Desirably, at least 95% by weight
of the particles of the active ingredient have an effective
particle size in the range of 0.01 to 10 micrometers.
[0061] A residue of a chemical species, as used in the
specification and concluding claims, refers to the moiety that is
the resulting product of the chemical species in a particular
reaction scheme or subsequent formulation or chemical product,
regardless of whether the moiety is actually obtained from the
chemical species. Thus, an ethylene glycol residue in a polymer
refers to one or more --OCH.sub.2CH.sub.2O-- units in the polter,
regardless of whether ethylene glycol was used to prepare the
polter. Similarly, a sebacic acid residue in a polter refers to one
or more --CO(CH.sub.2).sub.8CO-- moieties in the polter, regardless
of whether the residue is obtained by reacting sebacic acid or an
ester thereof to obtain the polymer. In certain aspects, a monomer
residue in a polymer can also be described as a repeating unit.
[0062] As used herein, the term "biologically active agent" or
"bioactive agent" means an agent that is capable of providing a
local or systemic biological, physiological, or therapeutic effect
in the biological system to which it is applied. For example, the
bioactive agent can act to control infection or inflammation,
enhance cell growth and tissue regeneration, control tumor growth,
act as an analgesic, promote anti-cell attachment, and enhance bone
growth, among other functions. Other suitable bioactive agents can
include anti-viral agents, vaccines, hormones, antibodies
(including active antibody fragments sFv, Fv, and Fab fragments),
aptamers, peptide mimetics, functional nucleic acids, therapeutic
proteins, peptides, or nucleic acids. Other bioactive agents
include prodrugs, which are agents that are not biologically active
when administered but, upon administration to a subject are
converted to bioactive agents through metabolism or some other
mechanism. Additionally, any of the compositions of the invention
can contain combinations of two or more bioactive agents. It is
understood that a biologically active agent can be used in
connection with administration to various subjects, for example, to
humans (i.e., medical administration) or to animals (i.e.,
veterinary administration).
[0063] As used herein, the term "pharmaceutically active agent"
includes a "drug" or a "vaccine" and means a molecule, group of
molecules, complex or substance administered to an organism for
diagnostic, therapeutic, preventative medical, or veterinary
purposes. This term include externally and internally administered
topical, localized and systemic human and animal pharmaceuticals,
treatments, remedies, nutraceuticals, cosmeceuticals, biologicals,
devices, diagnostics and contraceptives, including preparations
useful in clinical and veterinary screening, prevention,
prophylaxis, healing, wellness, detection, imaging, diagnosis,
therapy, surgery, monitoring, cosmetics, prosthetics, forensics and
the like. This term may also be used in reference to agriceutical,
workplace, military, industrial and environmental therapeutics or
remedies comprising selected molecules or selected nucleic acid
sequences capable of recognizing cellular receptors, membrane
receptors, hormone receptors, therapeutic receptors, microbes,
viruses or selected targets comprising or capable of contacting
plants, animals and/or humans. This term can also specifically
include nucleic acids and compounds comprising nucleic acids that
produce a bioactive effect, for example deoxyribonucleic acid (DNA)
or ribonucleic acid (RNA). Pharmaceutically active agents include
the herein disclosed categories and specific examples. It is not
intended that the category be limited by the specific examples.
Those of ordinary skill in the art will recognize also numerous
other compounds that fall within the categories and that are useful
according to the invention. Examples include a radiosensitizer, the
combination of a radiosensitizer and a chemotherapeutic, a steroid,
a xanthine, a beta-2-agonist bronchodilator, an anti-inflammatory
agent, an analgesic agent, a calcium antagonist, an
angiotensin-converting enzyme inhibitors, a beta-blocker, a
centrally active alpha-agonist, an alpha-1-antagonist, carbonic
anhydrase inhibitors, prostaglandin analogs, a combination of an
alpha agonist and a beta blocker, a combination of a carbonic
anhydrase inhibitor and a beta blocker, an
anticholinergic/antispasmodic agent, a vasopressin analogue, an
antiarrhythmic agent, an antiparkinsonian agent, an
antiangina/antihypertensive agent, an anticoagulant agent, an
antiplatelet agent, a sedative, an ansiolytic agent, a peptidic
agent, a biopolymeric agent, an antineoplastic agent, a laxative,
an antidiarrheal agent, an antimicrobial agent, an antifungal
agent, or a vaccine. In a further aspect, the pharmaceutically
active agent can be coumarin, albumin, bromolidine, steroids such
as betamethasone, dexamethasone, methylprednisolone, prednisolone,
prednisone, triamcinolone, budesonide, hydrocortisone, and
pharmaceutically acceptable hydrocortisone derivatives; xanthines
such as theophylline and doxophylline; beta-2-agonist
bronchodilators such as salbutamol, fenterol, clenbuterol,
bambuterol, salmeterol, fenoterol; antiinflammatory agents,
including antiasthmatic anti-inflammatory agents, antiarthritis
antiinflammatory agents, and non-steroidal antiinflammatory agents,
examples of which include but are not limited to sulfides,
mesalamine, budesonide, salazopyrin, diclofenac, pharmaceutically
acceptable diclofenac salts, nimesulide, naproxene, acetominophen,
ibuprofen, ketoprofen and piroxicam; analgesic agents such as
salicylates; calcium channel blockers such as nifedipine,
amlodipine, and nicardipine; angiotensin-converting enzyme
inhibitors such as captopril, benazepril hydrochloride, fosinopril
sodium, trandolapril, ramipril, lisinopril, enalapril, quinapril
hydrochloride, and moexipril hydrochloride; beta-blockers (i.e.,
beta adrenergic blocking agents) such as sotalol hydrochloride,
timolol maleate, timol hemihydrate, levobunolol hydrochloride,
esmolol hydrochloride, carteolol, propanolol hydrochloride,
betaxolol hydrochloride, penbutolol sulfate, metoprolol tartrate,
metoprolol succinate, acebutolol hydrochloride, atenolol, pindolol,
and bisoprolol fumarate; centrally active alpha-2-agonists (i.e.,
alpha adrenergic receptor agonist) such as clonidine, brimonidine
tartrate, and apraclonidine hydrochloride; alpha-1-antagonists such
as doxazosin and prazosin; anticholinergic/antispasmodic agents
such as dicyclomine hydrochloride, scopolamine hydrobromide,
glycopyrrolate, clidinium bromide, flavoxate, and oxybutynin;
vasopressin analogues such as vasopressin and desmopressin;
prostaglandin analogs such as latanoprost, travoprost, and
bimatoprost; cholinergics (i.e., acetylcholine receptor agonists)
such as pilocarpine hydrochloride and carbachol; glutamate receptor
agonists such as the N-methyl D-aspartate receptor agonist
memantine; anti-Vascular endothelial growth factor (VEGF) aptamers
such as pegaptanib; anti-VEGF antibodies (including but not limited
to anti-VEGF-A antibodies) such as ranibizumab and bevacizumab;
carbonic anhydrase inhibitors such as methazolamide, brinzolamide,
dorzolamide hydrochloride, and acetazolamide; antiarrhythmic agents
such as quinidine, lidocaine, tocamide hydrochloride, mexiletine
hydrochloride, digoxin, verapamil hydrochloride, propafenone
hydrochloride, flecaimide acetate, procainamide hydrochloride,
moricizine hydrochloride, and diisopyramide phosphate;
antiparkinsonian agents, such as dopamine, L-Dopa/Carbidopa,
selegiline, dihydroergocryptine, pergolide, lisuride, apomorphine,
and bromocryptine; antiangina agents and antihypertensive agents
such as isosorbide mononitrate, isosorbide dinitrate, propranolol,
atenolol and verapamil; anticoagulant and antiplatelet agents such
as coumadin, warfarin, acetylsalicylic acid, and ticlopidine;
sedatives such as benzodiazapines and barbiturates; ansiolytic
agents such as lorazepam, bromazepam, and diazepam; peptidic and
biopolymeric agents such as calcitonin, leuprolide and other LHRH
agonists, hirudin, cyclosporin, insulin, somatostatin, protirelin,
interferon, desmopressin, somatotropin, thymopentin, pidotimod,
erythropoietin, interleukins, melatonin,
granulocyte/macrophage-CSF, and heparin; antineoplastic agents such
as etoposide, etoposide phosphate, cyclophosphamide, methotrexate,
5-fluorouracil, vincristine, doxorubicin, cisplatin, hydroxyurea,
leucovorin calcium, tamoxifen, flutamide, asparaginase,
altretamine, mitotane, and procarbazine hydrochloride; laxatives
such as senna concentrate, casanthranol, bisacodyl, and sodium
picosulphate; antidiarrheal agents such as difenoxine
hydrochloride, loperamide hydrochloride, furazolidone,
diphenoxylate hydrochloride, and microorganisms; vaccines such as
bacterial and viral vaccines; antimicrobial agents such as
penicillins, cephalosporins, and macrolides, antifungal agents such
as imidazolic and triazolic derivatives; and nucleic acids such as
DNA sequences encoding for biological proteins, and antisense
oligonucleotides. It is understood that a pharmaceutically active
agent can be used in connection with administration to various
subjects, for example, to humans (i.e., medical administration) or
to animals (i.e., veterinary administration).
[0064] As used herein, the term "substituted" is contemplated to
include all permissible substituents of organic compounds. In a
broad aspect, the permissible substituents include acyclic and
cyclic, branched and unbranched, carbocyclic and heterocyclic, and
aromatic and nonaromatic substituents of organic compounds.
Illustrative substituents include, for example, those described
below. The permissible substituents can be one or more and the same
or different for appropriate organic compounds. For purposes of
this disclosure, the heteroatoms, such as nitrogen, can have
hydrogen substituents and/or any permissible substituents of
organic compounds described herein which satisfy the valences of
the heteroatoms. This disclosure is not intended to be limited in
any manner by the permissible substituents of organic compounds.
Also, the terms "substitution" or "substituted with" include the
implicit proviso that such substitution is in accordance with
permitted valence of the substituted atom and the substituent, and
that the substitution results in a stable compound, e.g., a
compound that does not spontaneously undergo transformation such as
by rearrangement, cyclization, elimination, etc. It is also
contemplated that, in certain aspects, unless expressly indicated
to the contrary, individual substituents can be further optionally
substituted (i.e., further substituted or unsubstituted).
[0065] In defining various terms, "A.sup.1," "A.sup.2," "A.sup.3,"
and "A.sup.4" are used herein as generic symbols to represent
various specific substituents. These symbols can be any
substituent, not limited to those disclosed herein, and when they
are defined to be certain substituents in one instance, they can,
in another instance, be defined as some other substituents.
[0066] The term "alkyl" as used herein is a branched or unbranched
saturated hydrocarbon group of 1 to 24 carbon atoms, such as
methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s-butyl,
t-butyl, n-pentyl, isopentyl, s-pentyl, neopentyl, hexyl, heptyl,
octyl, nonyl, decyl, dode cyl, tetradecyl, hexadecyl, eicosyl,
tetracosyl, and the like. The alkyl group can be cyclic or acyclic.
The alkyl group can be branched or unbranched. The alkyl group can
also be substituted or unsubstituted. For example, the alkyl group
can be substituted with one or more groups including, but not
limited to, alkyl, cycloalkyl, alkoxy, amino, ether, halide,
hydroxy, nitro, silyl, sulfo-oxo, or thiol, as described herein. A
"lower alkyl" group is an alkyl group containing from one to six
(e.g., from one to four) carbon atoms.
[0067] Throughout the specification "alkyl" is generally used to
refer to both unsubstituted alkyl groups and substituted alkyl
groups; however, substituted alkyl groups are also specifically
referred to herein by identifying the specific substituent(s) on
the alkyl group. For example, the term "halogenated alkyl" or
"haloalkyl" specifically refers to an alkyl group that is
substituted with one or more halide, e.g., fluorine, chlorine,
bromine, or iodine. The term "alkoxyalkyl" specifically refers to
an alkyl group that is substituted with one or more alkoxy groups,
as described below. The term "alkylamino" specifically refers to an
alkyl group that is substituted with one or more amino groups, as
described below, and the like. When "alkyl" is used in one instance
and a specific term such as "alkylalcohol" is used in another, it
is not meant to imply that the term "alkyl" does not also refer to
specific terms such as "alkylalcohol" and the like.
[0068] The term "alkynyl" as used herein is a hydrocarbon group of
2 to 24 carbon atoms with a structural formula containing at least
one carbon-carbon triple bond. The alkynyl group can be
unsubstituted or substituted with one or more groups including, but
not limited to, alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl,
alkynyl, cycloalkynyl, aryl, heteroaryl, aldehyde, amino,
carboxylic acid, ester, ether, halide, hydroxy, ketone, azide,
nitro, silyl, sulfo-oxo, or thiol, as described herein.
[0069] The terms "amine" or "amino" as used herein are represented
by the formula NA.sup.1A.sup.2, where A.sup.1 and A.sup.2 can be,
independently, hydrogen or alkyl, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group as
described herein.
[0070] The term "ester" as used herein is represented by the
formula --OC(O)A.sup.1 or --C(O)OA.sup.1, where A.sup.1 can be
alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl,
aryl, or heteroaryl group as described herein. The term "polter" as
used herein is represented by the formula
-(A.sup.1O(O)C-A.sup.2-C(O)O).sub.a-- or
-(A.sup.1O(O)C-A.sup.2-OC(O)).sub.a--, where A.sup.1 and A.sup.2
can be, independently, an alkyl, cycloalkyl, alkenyl, cycloalkenyl,
alkynyl, cycloalkynyl, aryl, or heteroaryl group described herein
and "a" is an integer from 1 to 500. "Polter" is as the term used
to describe a group that is produced by the reaction between a
compound having at least two carboxylic acid groups with a compound
having at least two hydroxyl groups.
[0071] The term "ether" as used herein is represented by the
formula A.sup.1OA.sup.2, where A.sup.1 and A.sup.2 can be,
independently, an alkyl, cycloalkyl, alkenyl, cycloalkenyl,
alkynyl, cycloalkynyl, aryl, or heteroaryl group described herein.
The term "polyether" as used herein is represented by the formula
-(A.sup.1O-A.sup.2O).sub.a--, where A.sup.1 and A.sup.2 can be,
independently, an alkyl, cycloalkyl, alkenyl, cycloalkenyl,
alkynyl, cycloalkynyl, aryl, or heteroaryl group described herein
and "a" is an integer of from 1 to 500. Examples of polyether
groups include polyethylene oxide, polypropylene oxide, and
polybutylene oxide.
[0072] The term "azide" as used herein is represented by the
formula --N.sub.3.
[0073] The term "thiol" as used herein is represented by the
formula --SH.
[0074] The terms "hydrolysable group" and "hydrolysable moiety"
refer to a functional group capable of undergoing hydrolysis, e.g.,
under basic or acidic conditions. Examples of hydrolysable residues
include, without limitation, acid halides, activated carboxylic
acids, and various protecting groups known in the art (see, for
example, "Protective Groups in Organic Synthesis," T. W. Greene, P.
G. M. Wuts, Wiley-Interscience, 1999).
[0075] Compounds described herein can contain one or more double
bonds and, thus, potentially give rise to cis/trans (E/Z) isomers,
as well as other conformational isomers. Unless stated to the
contrary, the invention includes all such possible isomers, as well
as mixtures of such isomers.
[0076] Unless stated to the contrary, a formula with chemical bonds
shown only as solid lines and not as wedges or dashed lines
contemplates each possible isomer, e.g., each enantiomer and
diastereomer, and a mixture of isomers, such as a racemic or
scalemic mixture. Compounds described herein can contain one or
more asymmetric centers and, thus, potentially give rise to
diastereomers and optical isomers. Unless stated to the contrary,
the present invention includes all such possible diastereomers as
well as their racemic mixtures, their substantially pure resolved
enantiomers, all possible geometric isomers, and pharmaceutically
acceptable salts thereof. Mixtures of stereoisomers, as well as
isolated specific stereoisomers, are also included. During the
course of the synthetic procedures used to prepare such compounds,
or in using racemization or epimerization procedures known to those
skilled in the art, the products of such procedures can be a
mixture of stereoisomers.
[0077] Certain materials, compounds, compositions, and components
disclosed herein can be obtained commercially or readily
synthesized using techniques generally known to those of skill in
the art. For example, the starting materials and reagents used in
preparing the disclosed compounds and compositions are either
available from commercial suppliers such as Aldrich Chemical Co.,
(Milwaukee, Wis.), Acros Organics (Morris Plains, N.J.), Fisher
Scientific (Pittsburgh, Pa.), or Sigma (St. Louis, Mo.) or are
prepared by methods known to those skilled in the art following
procedures set forth in references such as Fieser and Fieser's
Reagents for Organic Synthesis, Volumes 1-17 (John Wiley and Sons,
1991); Rodd's Chemistry of Carbon Compounds, Volumes 1-5 and
Supplementals (Elsevier Science Publishers, 1989); Organic
Reactions, Volumes 1-40 (John Wiley and Sons, 1991); March's
Advanced Organic Chemistry, (John Wiley and Sons, 4th Edition); and
Larock's Comprehensive Organic Transformations (VCH Publishers
Inc., 1989).
[0078] Unless otherwise expressly stated, it is in no way intended
that any method set forth herein be construed as requiring that its
steps be performed in a specific order. Accordingly, where a method
claim does not actually recite an order to be followed by its steps
or it is not otherwise specifically stated in the claims or
descriptions that the steps are to be limited to a specific order,
it is no way intended that an order be inferred, in any respect.
This holds for any possible non-express basis for interpretation,
including: matters of logic with respect to arrangement of steps or
operational flow; plain meaning derived from grammatical
organization or punctuation; and the number or type of embodiments
described in the specification.
[0079] Disclosed are the components to be used to prepare the
compositions of the invention as well as the compositions
themselves to be used within the methods disclosed herein. These
and other materials are disclosed herein, and it is understood that
when combinations, subsets, interactions, groups, etc. of these
materials are disclosed that while specific reference of each
various individual and collective combinations and permutation of
these compounds can not be explicitly disclosed, each is
specifically contemplated and described herein. For example, if a
particular compound is disclosed and discussed and a number of
modifications that can be made to a number of molecules including
the compounds are discussed, specifically contemplated is each and
every combination and permutation of the compound and the
modifications that are possible unless specifically indicated to
the contrary. Thus, if a class of molecules A, B, and C are
disclosed as well as a class of molecules D, E, and F and an
example of a combination molecule, A-D is disclosed, then even if
each is not individually recited each is individually and
collectively contemplated meaning combinations, A-E, A-F, B-D, B-E,
B-F, C-D, C-E, and C-F are considered disclosed. Likewise, any
subset or combination of these is also disclosed. Thus, for
example, the sub-group of A-E, B-F, and C-E would be considered
disclosed. This concept applies to all aspects of this application
including, but not limited to, steps in methods of making and using
the compositions of the invention. Thus, if there are a variety of
additional steps that can be performed it is understood that each
of these additional steps can be performed with any specific
embodiment or combination of embodiments of the methods of the
invention.
[0080] It is understood that the compositions disclosed herein have
certain functions. Disclosed herein are certain structural
requirements for performing the disclosed functions, and it is
understood that there are a variety of structures that can perform
the same function that are related to the disclosed structures, and
that these structures will typically achieve the same result.
B. POLYMERS
[0081] As briefly described herein, the present invention, in
various aspects, relates to glycidol-based polymer systems. In a
one aspect, as shown below, glycidol is an analog of ethylene
glycol. In further aspects, glycidol can be ring opened in
different ways, is capable of controlled polymerization, and is
inherently hydrophilic due to the presence of a primary hydroxyl
functionality. In one aspect, the glycidol polymers can be
semi-branched.
##STR00002##
[0082] In a further aspect, the semi-branched architectures can be
used for transportation of drugs and other biological cargo, as
shown in FIG. 1. However, this type of structure presents a number
of limitations. For example, the vastly branched systems have
limited post-modification, as they only contain an assortment of
primary and secondary hydroxyl groups, rather than an assortment of
reactive points. In a still further aspect, the random
configurations can lead to complications with introducing the
intended cargo to the system.
[0083] In further aspects, the glycidol based polymer systems
comprise linear glycidols. In one aspect, linear glycidols can be
accomplished using glycidol derivatives and anionic polymerization
methods, as represented by the reaction scheme below.
##STR00003##
[0084] However, this method involves rigorous reaction conditions,
is very susceptible to oxygen, and requires numerous purification
steps. In a further aspect, this method does not deliver polymer
systems with a suitable degradation profile. In a further aspect,
the inherent water solubility of poly(glycidol) systems can be
utilized to allow a method that will provide more linear
poly(glycidol) systems.
[0085] In a further aspect, the linear glycidols comprise
additional functional group containing co-monomers. In a still
further aspect, the additional functional groups can be
subsequently cross-linked to form nanoparticle structures. These
nanoparticles will imbibe the applicability of the water-soluble
glycidol units with the functionality of groups, such as allyl's
and epoxides, which are capable of a range of post-modification
reactions. In a further aspect, this will allow for the
encapsulation of drug molecules followed by the addition of
targeting units and/or dyes, as depicted in FIG. 2. In a still
further aspect, the addition of this increased functionality
provides a variety of nanoparticles that can be tailored to
specific needs, as well as structures whose functionality can be
verified in a laboratory setting. In an even further aspect, the
polymer systems can comprise polyester nanoparticle systems for the
transport of hydrophobic drug molecules.
[0086] As described above, glycidol's analogous structure to
polyethylene glycol, as well as its abundance of primary and
secondary hydroxyl groups provides a system that is relatively
non-toxic and exceedingly hydrophilic. In various aspects,
glycidol-based systems involve the formation of single-step
dendrimer like macromolecules that provide the abilities of
dendrimers without the painstaking process of dendrimer growth.[2,
4, 5, 18-22] In a further aspect, glycidol's success relates to its
latent AB.sub.2 monomer type. Thus, in a still further aspect, the
glycidol monomer does not become a true AB.sub.2 type monomer until
it has undergone ring opening. In various further aspects, this
characteristic allows for additional control through a ring-opening
polymerization rather than a rampant polycondensation, which is the
usual reaction type used with other AB.sub.2 type monomers.[4, 23,
24]
[0087] In a further aspect, the added control allows for
investigation of branched polyglycidols and the factors that lead
to branching. In a still further aspect, implementation of the
ring-opening polymerization (ROP) mechanism can yield materials
that are more chemically guided rather than empirical in the degree
of branching. In an even further aspect, hyperbranched polyglycidol
systems can be formed in varying sizes, with low polydispersity
indices (PDIs) and controlled degrees of polymerization (DP). In a
still further aspect, these polymer systems can be advantageous as
alternatives to multistep dendrimer species.[2, 4, 18-20, 22,
25]
[0088] In various aspects. the polymer systems formed from these
methods have use in numerous applications ranging from potential
vaccine models[26] and selective drug delivery vehicles[11-14, 27],
to biomineralization control and soluble catalyst supports in
organic synthesis. In a further aspect, much of the success seen in
these applications relates to the inherent characteristics of the
polyglycerol's branched structure. In a still further aspect,
limiting the degree of branching (DB), preferably with little
change to the PDI, can be beneficial in the formation of new, and
possibly more robust, poly(glycidol) architectures.
[0089] As briefly described, the current methods for the formation
of completely linear poly(glycidol) polymers rely on the use of
protected glycidol derivatives polymerized under stringent anionic
polymerization conditions. In a further aspect, the polymer then
undergoes a deprotection step that removes the protecting group,
leaving a linear poly(glycidol) structure. While an effective
method for the formation of linear poly(glycidol) species, this
method does not address the problems of the system.[28-31]
[0090] In some aspects, glycidol based polymers, much like
polyethylene glycol (PEG), have severely slow degradation profiles.
While not a significant problem for low molecular weight species,
the large macromolecular hyperbranched systems cannot be easily
eliminated from the body. In a further aspect, the difficult of
elimination from the body equates to an inevitable buildup of
poly(glycidol) (PG) over time and one would expect eventual data
will show higher toxicity with this build up, as is being seen now
with PEG. In a still further aspect, the presence of only one post
modification unit, the hydroxyl side arms, adds an extra challenge
to the creation of a polymer structure that has a variety of
post-modification capabilities.
[0091] In various further aspects, the present polyglycidol polymer
systems can be useful in the solubilization of proteins and siRNA,
which investigations had thus far been dominated by polyacrylates
having PEG side chains.[32, 33] In most aspects, RAFT initiators
are attached to thiol groups on the periphery of the structures,
and PEG is grown to cover the outside and increase
hydrophilicity.[34] Traditional methods are not ideal because they
severely diminish the activity of the protein as well as introduces
high molecular weight linear PEG into the body, which cannot easily
be eliminated.[35] It is this method of protein solubilization that
has been the cause of recent PEG toxicity problems. In some
aspects, the attachment of branched PEG systems to thiol-modified
siRNA and has shown an increase in biological half-life due to
reduced immunogenicity, arising from the morphology of the branched
structure, and enhanced resistance against proteolysis.[33]
[0092] In further aspects, poly(glycidol) can be used as a method
of increasing the solubility of biological structures, as depicted
in FIG. 3. In a still further aspect, the ability to control the
degree of branching present will be integral to the efficacy of the
synthesized structures and will allow for a more tailored approach
to solubilization of biological structures and their behavior in
vivo, as shown in FIGS. 4 and 5. In a yet further aspect,
facilitated by the increased degradability, the glycidol based
copolymers can be both more effective and less harmful than their
PEG counterparts.
[0093] Although a seemingly glaring problem, the low degree of
solubility in the polyglycerol systems has, for the most part, been
overlooked. Rather than attempting to form polyglycerols with
increased capabilities, a large amount of research has been aimed
at increasing the hydrophilicity of polyester structures. Using
polyether macroinitiators in the attempt to form block copolymers
is one method of combating this downfall of the polyester systems.
Unfortunately, the polymers synthesized in this manner are highly
prone to the formation of micellular structures, thus drastically
diminishing their actual viability.[1] Few studies have been
attempted on the basis of random copolymerization of glycidol with
other monomer species, and rarely address the branching
characteristics of the synthesized polymers.[24, 36] These
copolymerizations have also been dictated by the use of a single
catalyst, stannous ethylhexanoate, which is a common lactide
polymerization catalyst that implements a coordination-insertion
type mechanism.
[0094] The compounds and compositions described herein combat the
problems associated with polyglycerols, while still maintaining a
high degree of water solubility and low PDI values. In a further
aspect, the present disclosure provides inclusion of increased
physiological degradability, through the incorporation of esters,
as well as the introduction of more viable post modification units,
by the addition of allyl groups. In a further aspect, the
implementation of stannous triflate has been chosen as the desired
catalysis method based on its ability to allow for low reaction
temperatures while maintaining high polymerization rates and low
PDI values.
[0095] In a still further aspect, by employing the stannous
triflate catalyst at low temperatures, the DB of the resulting
polymers can be restricted to well below the currently published
values of 40% and higher. In still further aspect, the present
invention can comprise a range of comonomers, both commercially
available and novel, which exhibit the ability to copolymerize with
glycidol to form an array of new and exciting polymer
architectures. In a yet further aspect, the polymers comprise
desirable structural features and can provide new polymers with
customizable degrees of branching, high functionality, increased
solubility, and tunable biodegradability, thus imbuing all the
benefits of polyglycerols to systems that are more tailored for
delivery of a range of drugs and biological cargo. In an even
further aspect, the chemical characteristics of the synthesized
polymers were investigated through both .sup.1HNMR and .sup.13CNMR
techniques, and are further described in the Examples.
[0096] Disclosed herein is a polymer comprising repeating units
selected from:
##STR00004##
wherein R.sup.0 is selected from H, alkyl, NH.sub.2, and R.sup.1;
wherein R.sup.1 comprises a crosslinking functionality; wherein
repeating units A1, A2, B1, and B2 account for at least about 50
wgt % of the polymer; and wherein the ratio of (A1+A2):(B1+B2) is
greater than 1.
[0097] In one aspect, the ratio of (A1+A2):(B1+B2) is greater than
1. In another aspect, the ratio of (A1+A2):(B1+B2) is greater than
3. In yet another aspect, the ratio of (A1+A2):(B1+B2) is greater
than 5. In yet another aspect, the ratio of (A1+A2):(B1+B2) is
greater than 10. In yet another aspect, the ratio of
(A1+A2):(B1+B2) is greater than 25. In yet another aspect, the
ratio of (A1+A2):(B1+B2) is greater than 50. In yet another aspect,
the ratio of (A1+A2):(B1+B2) is greater than 100. In yet another
aspect, the ratio of (A1+A2):(B1+B2) is from 1 to 100. In yet
another aspect, the ratio of (A1+A2):(B1+B2) is from 5 to 100. In
yet another aspect, the ratio of (A1+A2):(B1+B2) is from 10 to 100.
In yet another aspect, the ratio of (A1+A2):(B1+B2) is from 25 to
100.
[0098] In one aspect, repeating units A1; A2; B1; and B2 account
for at least about 50 wgt % of the polymer. In another aspect,
repeating units A1; A2; B1; and B2 account for at least about 60
wgt % of the polymer. In yet another aspect, repeating units A1;
A2; B1; and B2 account for at least about 60 wgt % of the polymer.
In yet another aspect, repeating units A1; A2; B1; and B2 account
for at least about 70 wgt % of the polymer. In yet another aspect,
repeating units A1; A2; B1; and B2 account for at least about 80
wgt % of the polymer. In yet another aspect, repeating units A1;
A2; B1; and B2 account for at least about 90 wgt % of the polymer.
In yet another aspect, repeating units A1; A2; B1; and B2 account
for at least about 95 wgt % of the polymer. In yet another aspect,
repeating units A1; A2; B1; and B2 account for at least about 99
wgt % of the polymer.
[0099] In one aspect, the polymer is covalently bonded to a
biologic agent, such as a protein, DNA, or SiRNA, for example, a
protein. Such system can enhance the solubility of the biologic
agent.
[0100] In one aspect, the polymer comprising at least one repeating
unit formed from a monomer selected from:
##STR00005##
or a combination thereof.
[0101] In one aspect, the polymer comprises at least one repeating
unit from a monomer selected from:
##STR00006##
or a combination thereof, and wherein the polymer is oxidized to
form repeating units comprising epoxides or alkynes.
[0102] It is understood that all or only a portion of the repeating
units are oxidized in the polymer. Thus, it is understood that the
resultant polymer can comprise repeating units comprising alkenes
and repeating units comprising epoxides and/or alkynes. For
example, the polymer can comprise at least 1%, 5%, 10%, 15%, 20%,
or 25% repeating units that have been oxidized. Thus, in one
aspect, the polymer comprises repeating units comprising pendent
groups selected from
##STR00007##
or a combination thereof.
[0103] In one aspect, the polymer further comprises a repeating
unit formed from
##STR00008##
In another aspect, the polymer further comprises a repeating unit
formed from
##STR00009##
In yet another aspect, the polymer further comprises a repeating
unit formed from
##STR00010##
In yet another aspect, the polymer further comprises a repeating
unit formed from
##STR00011##
In yet another aspect, the polymer further comprises a repeating
unit formed from
##STR00012##
In yet another aspect, the polymer further comprises a repeating
unit formed from
##STR00013##
In yet another aspect, the polymer further comprises a repeating
unit formed from
##STR00014##
In yet another aspect, the polymer further comprises a repeating
unit formed from
##STR00015##
In yet another aspect, the polymer further comprises a repeating
unit formed from
##STR00016##
In yet another aspect, the polymer further comprises a repeating
unit formed from
##STR00017##
In yet another aspect, the polymer further comprises a repeating
unit formed from
##STR00018##
In yet another aspect, the polymer further comprises a repeating
unit formed from
##STR00019##
In yet another aspect, the polymer further comprises a repeating
unit formed from
##STR00020##
[0104] In one aspect, the polymer further comprises crosslinks,
wherein the crosslinks comprise
##STR00021##
wherein at least one of
##STR00022##
is not 0.
[0105] The crosslinks binds two or more polymers together. The
polymers can be any polymer disclosed herein. The crosslinks can
comprise one or more, such as two, moieties that can react with one
or more of the disclosed polymers thereby linking the polymers
together. Thus, suitable moieties include those that can react with
alkenes, epoxides, or alkynes. Non-limiting moieties include --SH,
--NH.sub.2, and
##STR00023##
The resultant polymer will then comprise one or more bonds which is
a result from these reactions. For example, the polymer can
comprise --S-- and --NH-- bonds. It is also understood that these
reactions will reduce the alkenes, alkynes, or epoxides that
participates in the reactions.
[0106] In one aspect, the polymer comprises
##STR00024## ##STR00025## ##STR00026## ##STR00027##
##STR00028##
[0107] Accordingly, the polymer can comprise the structure
##STR00029##
[0108] wherein Z.sup.1 is
##STR00030##
[0109] wherein Z.sup.2 is
##STR00031##
[0110] wherein Z.sup.3 is
##STR00032##
[0111] wherein Z.sup.4 is
##STR00033##
[0112] wherein Z.sup.5 is
##STR00034##
or a combination thereof; wherein, simultaneously, X.sub.1 is from
greater than 0% to 90%, X.sub.2 is from 0% to 95%, X.sub.3 is from
0% to 90%, X.sub.4 is from 0% to 90%, and X.sub.5 is from greater
than 0% to 90%, provided that
X.sub.1+X.sub.2+X.sub.3+X.sub.4+X.sub.5 equals 100%; and wherein
each R.sup.1 independently comprises a crosslinking
functionality
[0113] In one aspect, the crosslinking functionality comprises an
allyl, epoxide, amine, thiol, azide, or alkyne functionality. In
one aspect, the crosslinking functionality comprises an allyl
functionality. In another aspect, the crosslinking functionality
comprises an epoxide. In yet another aspect, the crosslinking
functionality comprises an amine functionality. In yet another
aspect, the crosslinking functionality comprises a thiol
functionality. In yet another aspect, the crosslinking
functionality comprises an azide functionality. In yet another
aspect, the crosslinking functionality comprises an alkyne
functionality.
[0114] In one aspect, polymer has the structure selected from the
group consisting of
##STR00035##
[0115] In one aspect, the polymer can comprise one or more of
repeating units selected from:
##STR00036##
or a combination thereof.
[0116] In another aspect, the polymer can comprise the
structure
##STR00037##
[0117] wherein each Z.sup.1 independently is
##STR00038##
[0118] wherein each Z.sup.2 independently is
##STR00039##
[0119] wherein Z.sup.3 is
##STR00040##
[0120] wherein Z.sup.4 is
##STR00041##
[0121] wherein each Z.sup.6 independently comprises
##STR00042##
and optionally independently comprises
##STR00043##
or a combination thereof; wherein, simultaneously, X.sub.1 is from
greater than 0% to 90%, X.sub.2 is from 0% to 95%, X.sub.3 is from
0% to 90%, X.sub.4 is from 0% to 90%, and X.sub.6 is from greater
than 0% to 90%, provided that
X.sub.1+X.sub.2+X.sub.3+X.sub.4+X.sub.6 equals 100%; wherein each
R.sup.1 independently comprises a crosslinking functionality; and
wherein L.sup.1 comprises
##STR00044##
wherein least one of
##STR00045##
is not 0.
[0122] In one aspect, the compound comprises the structure
##STR00046##
[0123] In one aspect, the polymer comprises a structure formed from
reacting a polymer disclosed herein further comprising repeating
units
##STR00047##
with a polymer comprising at least one repeating unit formed
from
##STR00048##
In one aspect, such polymer further comprises a structure formed
from reacting the polymer with a polymer comprising at least one
repeating unit formed from
##STR00049##
[0124] Also disclosed herein is a composition comprising a. a
polymer comprising repeating units formed from monomers
##STR00050##
wherein the polymer is crosslinked via crosslinks, wherein the
crosslinks comprises
##STR00051##
wherein at least one of
##STR00052##
is not 0; and b. a polymer comprising repeating units selected
from:
##STR00053##
wherein R.sup.0 is selected from H, alkyl, NH.sub.2, or R.sup.1;
wherein R.sup.1 comprises a crosslinking functionality; wherein
repeating units A1; A2; B1; and B2 account for at least about 50
wgt % of the polymer; and wherein the ratio of (A1+A2):(B1+B2) is
greater than 1, wherein the polymers are optionally covalently
bonded together.
[0125] Thus, the polymer can comprise the structure
##STR00054##
wherein L.sup.2 comprises
##STR00055##
wherein at least one of
##STR00056##
is not 0; and wherein each of x.sub.7, x.sub.8, x.sub.9, and
x.sub.10 independently are 1 to 1000. The polyglycidol is
reversibly attached and disconnected through transesterification
reactions. The attachment of the polyglycidol to the polycarbonate
via a transesterification reaction can be promoted by the use of
zinc acetate and heat. Such reaction is reversible. The attachment
and disconnection of the polyglycidol can be a function of the
temperature, thus, the structure of the compound can be controlled
by altering the temperature. Such compound is called a macroscopic
network and can be thermally responsive. Thus, the properties of
the compound can be altered by attaching or disconnecting the
polyglycidol to the polycarbonate polymer. A therapeutic agent,
diagnostic agent, or prophylactic agent, or a mixture thereof, can
be present in a composition comprising the compound or macroscopic
network. These macroscopic networks can be altered via reversible
trans-esterification reactions, thereby changing the properties of
the macroscopic network. An example of such technology is described
by Montarnal et al. (Science, 334, 965 (2011), which is hereby
incorporated by references in its entirety.
[0126] In one aspect, the polymer is a macroscopic network.
[0127] In one aspect, the polymer is biodegradable.
[0128] In one aspect, the polymer has a weight average molecular
weight from 1 kDa to 1,000 kDa. In another aspect, the polymer has
a weight average molecular weight from 1 kDa to 500 kDa. In yet
another aspect, the polymer has a weight average molecular weight
from 1 kDa to 150 kDa. In yet another aspect, the polymer has a
weight average molecular weight from 1 kDa to 100 kDa. In yet
another aspect, the polymer has a weight average molecular weight
from 1 kDa to 75 kDa. In yet another aspect, the polymer has a
weight average molecular weight from 1 kDa to 50 kDa. In yet
another aspect, the polymer has a weight average molecular weight
from 1 kDa to 25 kDa. In yet another aspect, the polymer has a
weight average molecular weight from 1 kDa to 10 kDa.
[0129] In one aspect, the polymer has a PDI from 1.01 to 5.0. In
another aspect, the polymer has a PDI from 1.01 to 4.0. In yet
another aspect, the polymer has a PDI from 1.01 to 3.0. In yet
another aspect, the polymer has a PDI from 1.01 to 2.0. In yet
another aspect, the polymer has a PDI from 1.01 to 1.5. In yet
another aspect, the polymer has a PDI from 1.01 to 1.25. In yet
another aspect, the polymer has a PDI from 1.01 to 1.10.
[0130] a. X Groups
[0131] In one aspect, X.sub.1+X.sub.2 equals from 50% to 99%. In
another aspect, X.sub.1+X.sub.2 equals from 60% to 95%. In yet
another aspect, X.sub.1+X.sub.2 equals from 70% to 90%.
[0132] In one aspect, X.sub.1+X.sub.2+X.sub.3+X.sub.4 equals from
50% to 99%. In another aspect, X.sub.1+X.sub.2+X.sub.3+X.sub.4
equals from 60% to 95%. In yet another aspect
X.sub.1+X.sub.2+X.sub.3+X.sub.4 equals from 70% to 90%.
[0133] In one aspect, X.sub.5 is from 1% to 40%. In another aspect,
X.sub.5 is from 5% to 35%. In yet another aspect, X.sub.5 is from
10% to 30%.
[0134] In one aspect, X.sub.2, X.sub.3, and X.sub.4 are 0%. In
another aspect, X.sub.2 is greater than 0% to 90% and X.sub.3, and
X.sub.4 are 0%. In yet another aspect, X.sub.2, X.sub.3, and
X.sub.4 are from greater than 0% to 90%.
[0135] In one aspect, each of X.sub.1, X.sub.2, X.sub.3, X.sub.4,
and X.sub.5 independently are 1 to 1000. In one aspect, each of
X.sub.1, X.sub.2, and X.sub.5 independently are 1 to 1000. In one
aspect, each of X.sub.1 and X.sub.5 independently are 1 to 1000. In
another aspect, each of X.sub.1, X.sub.2, X.sub.3, X.sub.4, and
X.sub.5 independently are 1 to 500. In yet another aspect, each of
X.sub.1, X.sub.2, X.sub.3, X.sub.4, and X.sub.5 independently are 1
to 300. In yet another aspect, each of X.sub.1, X.sub.2, X.sub.3,
X.sub.4, and X.sub.5 independently are 1 to 100. In yet another
aspect, each of X.sub.1, X.sub.2, X.sub.3, X.sub.4, and X.sub.5
independently are 1 to 50. In yet another aspect, each of X.sub.1,
X.sub.2, X.sub.3, X.sub.4, and X.sub.5 independently are 1 to 25.
In another aspect, each of X.sub.1, X.sub.2, and X.sub.5
independently are 1 to 500. In yet another aspect, each of X.sub.1,
X.sub.2, and X.sub.5 independently are 1 to 300. In yet another
aspect, each of X.sub.1, X.sub.2, and X.sub.5 independently are 1
to 100. In yet another aspect, each of X.sub.1, X.sub.2, and
X.sub.5 independently are 1 to 50. In yet another aspect, each of
X.sub.1, X.sub.2, and X.sub.5 independently are 1 to 25. In another
aspect, each of X.sub.1 and X.sub.5 independently are 1 to 500. In
yet another aspect, each of X.sub.1 and X.sub.5 independently are 1
to 300. In yet another aspect, each of X.sub.1 and X.sub.5
independently are 1 to 100. In yet another aspect, each of X.sub.1
and X.sub.5 independently are 1 to 50. In yet another aspect, each
of X.sub.1 and X.sub.5 independently are 1 to 25.
[0136] In one aspect, each of X.sub.1, X.sub.2, X.sub.3, X.sub.4,
and X.sub.6 independently are 1 to 1000. In one aspect, each of
X.sub.1, X.sub.2, and X.sub.6 independently are 1 to 1000. In one
aspect, each of X.sub.1 and X.sub.6 independently are 1 to 1000. In
another aspect, each of X.sub.1, X.sub.2, X.sub.3, X.sub.4, and
X.sub.6 independently are 1 to 500. In yet another aspect, each of
X.sub.1, X.sub.2, X.sub.3, X.sub.4, and X.sub.6 independently are 1
to 300. In yet another aspect, each of X.sub.1, X.sub.2, X.sub.3,
X.sub.4, and X.sub.6 independently are 1 to 100. In yet another
aspect, each of X.sub.1, X.sub.2, X.sub.3, X.sub.4, and X.sub.6
independently are 1 to 50. In yet another aspect, each of X.sub.1,
X.sub.2, X.sub.3, X.sub.4, and X.sub.6 independently are 1 to 25.
In another aspect, each of X.sub.1, X.sub.2, and X.sub.6
independently are 1 to 500. In yet another aspect, each of X.sub.1,
X.sub.2, and X.sub.6 independently are 1 to 300. In yet another
aspect, each of X.sub.1, X.sub.2, and X.sub.6 independently are 1
to 100. In yet another aspect, each of X.sub.1, X.sub.2, and
X.sub.6 independently are 1 to 50. In yet another aspect, each of
X.sub.1, X.sub.2, and X.sub.6 independently are 1 to 25. In another
aspect, each of X.sub.1 and X.sub.6 independently are 1 to 500. In
yet another aspect, each of X.sub.1 and X.sub.6 independently are 1
to 300. In yet another aspect, each of X.sub.1 and X.sub.6
independently are 1 to 100. In yet another aspect, each of X.sub.1
and X.sub.6 independently are 1 to 50. In yet another aspect, each
of X.sub.1 and X.sub.6 independently are 1 to 25.
[0137] In one aspect, each of x.sub.7, x.sub.8, x.sub.9, and
x.sub.10 independently are 1 to 1000. In another aspect, each of
x.sub.7, x.sub.8, x.sub.9, and x.sub.10 independently are 1 to 500.
In yet another aspect, each of x.sub.7, x.sub.8, x.sub.9, and
x.sub.10 independently are 1 to 300. In yet another aspect, each of
x.sub.7, x.sub.8, x.sub.9, and x.sub.10 independently are 1 to 100.
In yet another aspect, each of x.sub.7, x.sub.8, x.sub.9, and
x.sub.10 independently are 1 to 50. In yet another aspect, each of
x.sub.7, x.sub.8, x.sub.9, and x.sub.10 independently are 1 to
25.
b. Z Groups
[0138] It is understood that the Z groups represent It is
understood that the arrangement of Z groups in the polymers
disclosed herein can be in any order, for example, Z.sup.1,
Z.sup.2, Z.sup.3, Z.sup.4, and Z.sup.5 can be in any order. Thus,
it is also understood that the polymer can be a random copolymer,
whereby the order of each repeat unit of Z.sup.1, Z.sup.2, Z.sup.3,
Z.sup.4, and Z.sup.5 is random.
[0139] In one aspect, Z.sup.1 is
##STR00057##
In another aspect, Z.sup.1 is
##STR00058##
In yet another aspect, Z.sup.1 is
##STR00059##
In yet another aspect, Z.sup.1 is
##STR00060##
[0140] In one aspect, Z.sup.2 is
##STR00061##
In another aspect, Z.sup.2 is
##STR00062##
[0141] In one aspect, Z.sup.5 is
##STR00063##
In another aspect, Z.sup.5 is
##STR00064##
In yet another aspect, Z.sup.5 is
##STR00065##
In yet another aspect, Z.sup.5 is
##STR00066##
[0142] In yet another aspect, Z.sup.5 is
##STR00067##
In yet another aspect, Z.sup.5 is
##STR00068##
In yet another aspect, Z.sup.5 is
##STR00069##
[0143] In one aspect, each Z.sup.6 comprises
##STR00070##
In another aspect, each Z.sup.6 comprises
##STR00071##
In one aspect, Z.sup.11 is
##STR00072##
In one aspect, Z.sup.12 is
##STR00073##
In one aspect, Z.sup.13 is
##STR00074##
c. R Groups
[0144] In one aspect, R.sup.0 is H. In another aspect, R.sup.0 is
alkyl. In yet another aspect, R.sup.0 is NH.sub.2. In yet another
aspect, R.sup.0 is R.sup.1.
[0145] In one aspect, each of R.sup.1 and R.sup.2 independently
comprises
##STR00075##
wherein at least one of
##STR00076##
is not 0.
[0146] In one aspect, R.sup.1 comprises
##STR00077##
for example, R.sup.1 can comprise
##STR00078##
In one aspect, R.sup.1 comprises
##STR00079##
In another aspect, R.sup.1 comprises
##STR00080##
In one aspect, R.sup.1 comprises
##STR00081##
In yet another aspect, R.sup.1 comprises
##STR00082##
In yet another aspect, R.sup.1 comprises
##STR00083##
and an allyl functionality.
[0147] In one aspect, R.sup.2 comprises
##STR00084##
for example, R.sup.2 can comprise
##STR00085##
In one aspect, R.sup.2 comprises
##STR00086##
In another aspect, R.sup.2 comprises
##STR00087##
In one aspect, R.sup.2 comprises
##STR00088##
In yet another aspect, R.sup.2 comprises
##STR00089##
In yet another aspect, R.sup.1 comprises
##STR00090##
and an allyl functionality.
[0148] In one aspect, the each of R.sup.1 and R.sup.2 independently
are selected from the group consisting of
##STR00091##
[0149] In one aspect, R.sup.1 is
##STR00092##
In another aspect, R.sup.1 is
##STR00093##
In yet another aspect, R.sup.1 is
##STR00094##
In yet another aspect, R.sup.1 is
##STR00095##
In yet another aspect, R.sup.1 is
##STR00096##
In yet another aspect, R.sup.1 is
##STR00097##
In yet another aspect, R.sup.1 is
##STR00098##
In yet another aspect, R.sup.1 is
##STR00099##
In yet another aspect, R.sup.1 is
##STR00100##
In yet another aspect, R.sup.1 is
##STR00101##
In yet another aspect, R.sup.1 is
##STR00102##
[0150] In one aspect, R.sup.2 is
##STR00103##
In another aspect, R.sup.2 is
##STR00104##
In yet another aspect, R.sup.2 is
##STR00105##
In yet another aspect, R.sup.2 is
##STR00106##
In yet another aspect, R.sup.2 is
##STR00107##
In yet another aspect, R.sup.2 is
##STR00108##
In yet another aspect, R.sup.2 is
##STR00109##
d. Crosslinks and L Groups
[0151] In one aspect, crosslinks or L.sup.1 comprises
##STR00110##
wherein least one of
##STR00111##
is not 0.
[0152] In one aspect, crosslinks or L.sup.1 comprises at least
##STR00112##
In one aspect, L.sup.1 comprises at least
##STR00113##
[0153] In one aspect, crosslinks or L.sup.1 comprises one or more
of
##STR00114##
or any combination thereof.
[0154] In one aspect, crosslinks or L.sup.1 comprises one or more
of
##STR00115##
In another aspect, crosslinks or L.sup.1 comprises one or more
of
##STR00116##
In yet another aspect, crosslinks or L.sup.1 comprises one or more
of
##STR00117##
In yet another aspect, crosslinks or L.sup.1 comprises one or more
of
##STR00118##
In yet another aspect, crosslinks or L.sup.1 comprises one or more
of
##STR00119##
[0155] In one aspect, L.sup.1 comprises one or more of
##STR00120##
or any combination thereof.
[0156] In one aspect, L.sup.1 comprises two of
##STR00121##
or any combination thereof.
[0157] In another aspect, L.sup.1 comprises two of
##STR00122##
or any combination thereof.
[0158] In one aspect, L.sup.1 is
##STR00123## ##STR00124## ##STR00125## ##STR00126##
##STR00127##
[0159] In one aspect, L.sup.2 comprises
##STR00128##
wherein at least one of
##STR00129##
is not 0.
[0160] In one aspect, L.sup.2 comprises at least
##STR00130##
In one aspect, L.sup.2 comprises at least
##STR00131##
[0161] In one aspect, L.sup.2 comprises
##STR00132##
or any combination thereof.
[0162] In a further aspect, the invention relates to a polyglycidol
having a degree of branching of less than about 0.25. For example,
the degree of branching can less than about 0.20, less than about
0.15, or less than about 0.10.
C. NANOPARTICLES
[0163] Also disclosed herein is a nanoparticle comprising one or
more compounds or polymers disclosed herein. In one aspect, the
nanoparticle is made from one or more compounds or polymers
disclosed herein, for example, the nanoparticle is made from one or
more polymers disclosed herein. In one aspect, the nanoparticle
comprises crosslinked polymers disclosed herein.
[0164] In one aspect, the nanoparticle further comprises at least
one pharmaceutically active agent and/or biologically active
agent.
[0165] In one aspect, the nanoparticle is biodegradable. The
biodegradability can depend on the number of hydrolysable bonds,
such as ester bonds, present in the compounds making up the
nanoparticle.
[0166] In one aspect, the nanoparticle is hydrophilic. In another
aspect, the t at least one pharmaceutically active agent and/or
biologically active agent is hydrophobic. In another aspect, the at
least one pharmaceutically active agent and/or biologically active
agent is a protein, DNA, or SiRNA. In one aspect, the at least one
pharmaceutically active agent and/or biologically active agent is
covalently bonded to the nanoparticle.
[0167] In one aspect, the nanoparticle is between 1 nm and 1000 nm
in diameter. In another aspect, the nanoparticle is between 1 nm
and 750 nm in diameter. In yet another aspect, the nanoparticle is
between 1 nm and 500 nm in diameter. In yet another aspect, the
nanoparticle is between 1 nm and 250 nm in diameter. In yet another
aspect, the nanoparticle is between 1 nm and 100 nm in
diameter.
[0168] In one aspect, the nanoparticle comprises reactive
functionalities, such as a hydroxyl group, an amine group, a thiol
group, an allyl group, an epoxide, or an alkyne group, or a
combination thereof.
D. COMPOSITIONS AND PHARMACEUTICAL COMPOSITIONS
[0169] Also disclosed herein are compositions, such as
pharmaceutical compositions.
[0170] In one aspect, the pharmaceutical composition comprises a) a
compound or polymer disclosed herein; b) pharmaceutically active
agent and/or biologically active agent; and c) a pharmaceutically
acceptable carrier.
[0171] In one aspect, the pharmaceutical composition comprises a) a
nanoparticle disclosed herein; b) pharmaceutically active agent
and/or biologically active agent; and c) a pharmaceutically
acceptable carrier.
[0172] In one aspect, the invention relates to pharmaceutical
compositions comprising the disclosed compounds and/or
nanoparticles; pharmaceutically active agent and/or biologically
active agent, and a pharmaceutically acceptable carrier or salt
thereof. In an aspect, the disclosed pharmaceutical compositions
can be provided comprising a therapeutically effective amount of
the therapeutic agent, diagnostic agent, or prophylactic agent, or
a mixture thereof, and a pharmaceutically acceptable carrier. The
disclosed pharmaceutical compositions can be provided comprising a
prophylactically effective amount of the therapeutic agent,
diagnostic agent, or prophylactic agent, or a mixture thereof, and
pharmaceutically acceptable carrier.
[0173] In one aspect, the pharmaceutical composition comprises one
or more pharmaceutically active agent and/or biologically active
agents. The compounds and nanoparticles disclosed herein are
capable of being loaded with several different classes of
therapeutics. Thus, the pharmaceutical composition is capable of
delivering at least two different classes of therapeutics. For
example, the therapeutic agent can comprise a MEK inhibitor and a
bone morphogenetic protein 2 (BMP2) growth factor. In one aspect,
the therapeutic agent is hydrophobic. The compounds and
nanoparticles are capable of being a delivery vehicle for
therapeutic agents, diagnostic agents, or prophylactic agents that
were previously difficult to deliver due to their physical
properties, such as their hydrophobicity. In one aspect, an
effective amount of a therapeutic agent, diagnostic agent, or
prophylactic agent can be present in the pharmaceutical
composition. For example, an effective amount of a therapeutic
agent, diagnostic agent, or prophylactic agent can be loaded in the
compounds or nanoparticles disclosed herein.
[0174] The pharmaceutical carrier employed can be, for example, a
solid, liquid, or gas. Examples of solid carriers include lactose,
terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium
stearate, and stearic acid. Examples of liquid carriers are sugar
syrup, peanut oil, olive oil, and water. Examples of gaseous
carriers include carbon dioxide and nitrogen.
[0175] In preparing the compositions for oral dosage form, any
convenient pharmaceutical media can be employed. For example,
water, glycols, oils, alcohols, flavoring agents, preservatives,
coloring agents and the like can be used to form oral liquid
preparations such as suspensions, elixirs and solutions; while
carriers such as starches, sugars, microcrystalline cellulose,
diluents, granulating agents, lubricants, binders, disintegrating
agents, and the like can be used to form oral solid preparations
such as powders, capsules and tablets. Because of their ease of
administration, tablets and capsules are the preferred oral dosage
units whereby solid pharmaceutical carriers are employed.
Optionally, tablets can be coated by standard aqueous or nonaqueous
techniques
[0176] The instant compositions include compositions suitable for
oral, rectal, topical, and parenteral (including subcutaneous,
intramuscular, and intravenous) administration, although the most
suitable route in any given case will depend on the particular
host, and nature and severity of the conditions for which the
active ingredient is being administered. The pharmaceutical
compositions can be conveniently presented in unit dosage form and
prepared by any of the methods well known in the art of
pharmacy.
[0177] Pharmaceutical compositions of the present invention
suitable for parenteral administration can be prepared as solutions
or suspensions of the active compounds in water. A suitable
surfactant can be included such as, for example,
hydroxypropylcellulose. Dispersions can also be prepared in
glycerol, liquid polyethylene glycols, and mixtures thereof in
oils. Further, a preservative can be included to prevent the
detrimental growth of microorganisms.
[0178] Pharmaceutical compositions of the present invention
suitable for injectable use include sterile aqueous solutions or
dispersions. Furthermore, the compositions can be in the form of
sterile powders for the extemporaneous preparation of such sterile
injectable solutions or dispersions. In all cases, the final
injectable form must be sterile and must be effectively fluid for
easy syringability. The pharmaceutical compositions must be stable
under the conditions of manufacture and storage; thus, preferably
should be preserved against the contaminating action of
microorganisms such as bacteria and fungi. The carrier can be a
solvent or dispersion medium containing, for example, water,
ethanol, polyol (e.g., glycerol, propylene glycol and liquid
polyethylene glycol), vegetable oils, and suitable mixtures
thereof.
[0179] Pharmaceutical compositions of the present invention can be
in a form suitable for topical use such as, for example, an
aerosol, cream, ointment, lotion, dusting powder, mouth washes,
gargles, and the like. Further, the compositions can be in a form
suitable for use in transdermal devices. These formulations can be
prepared, utilizing a compound of the invention, or
pharmaceutically acceptable salts thereof, via conventional
processing methods. As an example, a cream or ointment is prepared
by mixing hydrophilic material and water, together with about 5 wt
% to about 10 wt % of the compound, to produce a cream or ointment
having a desired consistency.
[0180] Pharmaceutical compositions of this invention can be in a
form suitable for rectal administration wherein the carrier is a
solid. It is preferable that the mixture forms unit dose
suppositories. Suitable carriers include cocoa butter and other
materials commonly used in the art. The suppositories can be
conveniently formed by first admixing the composition with the
softened or melted carrier(s) followed by chilling and shaping in
moulds.
[0181] In addition to the aforementioned carrier ingredients, the
pharmaceutical formulations described above can include, as
appropriate, one or more additional carrier ingredients such as
diluents, buffers, flavoring agents, binders, surface-active
agents, thickeners, lubricants, preservatives (including
anti-oxidants) and the like. Furthermore, other adjuvants can be
included to render the formulation isotonic with the blood of the
intended recipient. Compositions containing a compound of the
invention, and/or pharmaceutically acceptable salts thereof, can
also be prepared in powder or liquid concentrate form.
[0182] It is understood, however, that the specific dose level for
any particular patient will depend upon a variety of factors. Such
factors include the age, body weight, general health, sex, and diet
of the patient. Other factors include the time and route of
administration, rate of excretion, drug combination, and the type
and severity of the particular disease undergoing therapy.
[0183] The present invention is further directed to a method for
the manufacture of a medicament for treatment of a disorder in a
subject (e.g., humans) comprising combining one or more disclosed
compounds, products, or compositions with a pharmaceutically
acceptable carrier or diluent. Thus, in one aspect, the invention
relates to a method for manufacturing a medicament comprising
combining at least one disclosed compound, a therapeutic agent,
diagnostic agent, or prophylactic agent, or a mixture thereof, with
a pharmaceutically acceptable carrier or diluent.
[0184] It is understood that the disclosed compositions can be
prepared from the disclosed compounds. It is also understood that
the disclosed compositions can be employed in the disclosed methods
of using.
[0185] a. Methods of Using the Pharmaceutical Compositions
[0186] Disclosed herein is a drug delivery method comprising the
step of administering to a subject a composition comprising a
polymer or nanoparticle disclosed herein, in combination with at
least one pharmaceutically active agent and/or biologically active
agent. In one aspect, the composition further comprises a
pharmaceutically acceptable carrier. In one aspect, the method
comprises administering an effective amount of the pharmaceutically
active agent and/or biologically active agent to the subject. In
one aspect, the effective amount is a therapeutically effective
amount. Such amount can be determined by one skilled in the
art.
[0187] In one aspect, the therapeutic agent is a cancer agent. In
another aspect, the therapeutic agent is a protein, DNA, or
SiRNA.
[0188] In one aspect, the subject is an animal. In a further
aspect, the subject is a mammal. In a yet further aspect, the
subject is a primate. In a still further aspect, the subject is a
human. In an even further aspect, the subject is a patient.
[0189] In a further aspect, the pharmaceutical composition is
administered following identification of the subject in need of
treatment of disorder. In a still further aspect, the
pharmaceutical composition is administered following identification
of the subject in need of prevention of a disorder. In an even
further aspect, the subject has been diagnosed with a need for
treatment of a disorder prior to the administering step.
[0190] In one aspect, the method delivers one or more therapeutic
agents. The compounds and nanoparticles disclosed herein are
capable of being loaded with several different classes of
therapeutics. Thus, the method is capable of delivering at least
two different classes of therapeutics. For example, the therapeutic
agent can comprise a MEK inhibitor and a bone morphogenetic protein
2 (BMP2) growth factor.
[0191] In one aspect, the method comprises administering an
effective amount of the pharmaceutically active agent and/or
biologically active agent to the subject. In one aspect, the
effective amount is a therapeutically effective amount. Such amount
can be determined by one skilled in the art.
[0192] In one aspect, the therapeutic agent is a cancer agent. In
another aspect, the pharmaceutically active agent and/or
biologically active agent is a protein, DNA, or SiRNA.
[0193] In one aspect, the subject is an animal. In a further
aspect, the subject is a mammal. In a yet further aspect, the
subject is a primate. In a still further aspect, the subject is a
human. In an even further aspect, the subject is a patient.
[0194] In a further aspect, the pharmaceutical composition is
administered following identification of the subject in need of
treatment of disorder. In a still further aspect, the
pharmaceutical composition is administered following identification
of the subject in need of prevention of a disorder. In an even
further aspect, the subject has been diagnosed with a need for
treatment of a disorder prior to the administering step.
E. METHOD OF MAKING POLYMERS
[0195] Also disclosed here is a method of making a polymer, the
method comprising the step of polymerizing glycidol in the presence
of a tin catalyst. In one aspect, the tin catalyst is a tin(II)
catalyst, for example, Sn(OTf).sub.2.
[0196] Also disclosed is a polymer made from the methods disclosed
herein. For example, the resultant polymer comprises repeating
units selected from:
##STR00133##
wherein R.sup.0 is selected from H, alkyl, NH.sub.2, and R.sup.1;
wherein R.sup.1 comprises a crosslinking functionality; wherein
repeating units A1, A2, B1, and B2 account for at least about 50
wgt % of the polymer; and wherein the ratio of (A1+A2):(B1+B2) is
greater than 1.
[0197] In another aspect, the resultant polymer further comprises
at least one repeating unit formed from a monomer selected
from:
##STR00134##
or a combination thereof.
[0198] In one aspect, the method further comprises the step of
crosslinking the polymer with crosslinks, wherein the wherein the
crosslinks comprises
##STR00135##
wherein at least one of
##STR00136##
is not 0.
[0199] In another aspect, the polymer is linear when the first
compound comprises an ester moiety. In another aspect, the polymer
is semi-branched when the first polymer comprises a glycidol
moiety. Non-limiting examples of first compounds comprising an
ester moiety are 2-oxepane-1,5-dione and lactones, such as
.delta.-valerolactone and .alpha.-allyl-.delta.-valerolactone.
Non-limiting examples of first compounds comprising a glycidol
moiety are glycidol, allyl-glycidol ether, glycidyl ester allyl,
and ethoxyethyl glycidyl ether. In one aspect, the method comprises
polymerizing the first compound comprising a glycidol moiety and/or
an ester moiety with a second compound comprising a glycidol moiety
and/or a ester moiety, thereby making a copolymer. In one aspect,
the first compound comprises a glycidol moiety and the second
compound comprises an ester moiety. In one aspect, the first
compound comprises a glycidol moiety and the second compound
comprises a glycidol moiety and an ester moiety.
[0200] In one aspect, the polymerization step is performed at a
temperature of from -30.degree. C. to 50.degree. C. In another
aspect, the polymerization step is performed at a temperature of
from -30.degree. C. to 20.degree. C. In yet another aspect, the
polymerization step is performed at a temperature of from
-30.degree. C. to 0.degree. C. In yet another aspect, the
polymerization step is performed at a temperature of from
-30.degree. C. to -10.degree. C.
[0201] Also disclosed herein is a method of crosslinking
comprising: a) providing a first and second compound, wherein both
the first and second compound comprises a crosslinking
functionality; b) crosslinking the first and second compound with a
crosslinks described herein.
F. MANUFACTURE OF A MEDICAMENT
[0202] In one aspect, the invention relates to a method for the
manufacture of a medicament for treatment of a disorder comprising
combining a disclosed compound or nanoparticle with a
therapeutically effective amount of a therapeutic agent, diagnostic
agent, or prophylactic agent, or a mixture thereof and with a
pharmaceutically acceptable carrier or diluent.
G. KITS
[0203] Disclosed herein is a kit comprising a compound or
nanoparticle disclosed herein and a therapeutic agent, diagnostic
agent, or prophylactic agent, or a mixture thereof and one or more
of: a) instructions of delivering the therapeutic agent, diagnostic
agent, or prophylactic agent, or a mixture thereof; b) instructions
for using the therapeutic agent, diagnostic agent, or prophylactic
agent, or a mixture thereof to treat a disorder.
H. EXPERIMENTAL
[0204] The following examples are put forth so as to provide those
of ordinary skill in the art with a complete disclosure and
description of how the compounds, compositions, articles, devices
and/or methods claimed herein are made and evaluated, and are
intended to be purely exemplary of the invention and are not
intended to limit the scope of what the inventors regard as their
invention. However, those of skill in the art should, in light of
the present disclosure, appreciate that many changes can be made in
the specific embodiments which are disclosed and still obtain a
like or similar result without departing from the spirit and scope
of the invention.
[0205] Efforts have been made to ensure accuracy with respect to
numbers (e.g., amounts, temperature, etc.), but some errors and
deviations should be accounted for. Unless indicated otherwise,
parts are parts by weight, temperature is in .degree. C. or is at
ambient temperature, and pressure is at or near atmospheric.
[0206] As briefly discussed above, while hyperbranched systems
formed through the polymerization of glycidol have shown
applicability, the ability to form polymers with a controlled
degree of branching was investigated. In one aspect, controlled
polymerization can allow the lower branching systems to achieve a
better clearance but also allow the formation of nanoparticles
which is not possible with globular starticel materials. The
semibraching retain some benefits of the hyperbranched systems for
the functionalization and hydrogels formation with and without
stimuli-responsive reactions (FIG. 27-28). Without wishing to be
bound by a particular theory, it is believed that the secondary
reaction ability will impart a wider range of versatility to the
already robust poly(glycidol) architecture. In a further aspect,
the increase in post-modification capability will increase the
viability of the synthesized polymer systems and allow for novel
macromolecules. As depicted in FIG. 6, introduction of allyl
functionalities, in one aspect, can allow for the formation of a
more robust polymer system with crosslinking potential.
[0207] In a still further aspect, controlled degree of branching
and linear systems which can be synthesized more easily can used
for targeted delivery of drug molecules and biological cargo. In a
yet further aspect, semi-branched structures with that are more
favorable in vivo.
[0208] In various aspects, the ring opening polymerization of
glycidol can be influenced kinetically. In the present example, the
kinetic control on the polymerization of poly(glycidol) was
evaluated. To evaluate the kinetics of the polymer system, four
temperatures were chosen in order to undergo a thorough kinetic
study on the ring opening polymerization mechanism. The
temperatures chosen are shown in Table 1, below. The degree of
branching in the resultant polymers can be calculated using
equation 1 shown below, with the variables referring to the
integration values obtained from quantitative .sup.13CNMR
investigation, as further shown in FIG. 7. Unique peaks arise in
the .sup.13C-NMR based on the type of ring opening undergone by
each monomer. The .sup.13C-NMR of glycidol homopolymer is shown in
FIG. 8.
DB = 2 D 2 D + L 1 , 3 + L 1 , 4 ( Eq . 1 ) ##EQU00001##
[0209] For the purpose of direct correlation, the only variable
that was changed for each reaction was the temperature. NMR data
and degree of branching for polyglycidol systems are reported in
Table 1 below. As shown in Table 1 and FIG. 9, kinetic control over
the degree of branching in the polymer systems was accomplished by
depressing the temperature at which the reaction was conducted.
Inspection of the NMR data (FIG. 10) shows the suppression of the
dendritic carbon peak while the linear peak remains strong. Thus,
depressed reaction temperatures reduce the formation of branches
within the polymer backbone. In a further aspect, these results
allows for the determination of an optimal reaction temperature
based on the degree of branching that is desired for the various
applications proposed for the synthesized polymers.
TABLE-US-00001 TABLE 1 Glycidol Homopolymer Reaction Temperature
(.degree. C.) Region Shift (ppm) 40.degree. C. 20.degree. C.
0.degree. C. -20.degree. C. L.sub.1, 3 81.0-82.0 1.00 1.00 1.00
1.00 D 79.5-80.5 0.79 0.62 0.60 0.48 2 L.sub.1, 4 73.5-74.5 3.69
3.65 3.98 4.80 2D, 2T 72.0-73.5 7.05 7.52 7.62 8.44 L.sub.1, 3,
L.sub.1, 4 70.5-72.0 3.14 3.01 3.17 2.93 T 64.0-65.0 1.74 2.13 2.17
3.53 L.sub.1, 3 62.0-63.5 3.15 2.97 2.76 2.87 Degree of Branching
0.24 0.21 0.20 0.15 Relative Abundance of 10.5% 8.2% 8.0% 5.2%
Dendritic Carbons Glycidol Homopolymer Temperature (.degree. C.)
Region Shift (ppm) 25.degree. C. -5.degree. C. -42.degree. C.
-78.degree. C. L.sub.1, 3 81.0-82.0 1.00 1.00 1.00 1.00 D 79.5-80.5
0.78 0.41 0.35 0 2 L.sub.1, 4 73.5-74.5 3.49 4.57 5.14 5.58 2D, 2T
72.0-73.5 7.55 8.99 7.54 8.42 L.sub.1, 3, L.sub.1, 4 70.5-72.0 3.91
4.05 2.92 2.67 T 64.0-65.0 1.71 3.60 4.10 8.10 L.sub.1, 3 62.0-63.5
3.07 3.34 2.86 3.69 Degree of Branching 0.2447 0.1272 0.1142 0
[0210] The capability to choose the degree of branching optimized
for the poly(glycidol) system affords the ability to modify the
synthesized polymers based on the preferred application for each.
This possibility indicates that the present invention can be used
in a wide range of applications with the reaction temperature being
the determining factor for the polymer architecture. Without
wishing to be bound by a particular theory, such control over this
synthetic method allows for more effective and diverse potential.
Poly(glycidol) branching possibilities are shown in FIGS. 11 and
12.
[0211] In various aspects, the glycidol monomer can be opened in
two ways to yield different polymer units.[4, 22, 24, 36] As seen
in FIG. 13, these repeat units are known as linear-1,3 (L.sub.1,3)
and linear-1,4 (L.sub.1,4) and influence the branching that is seen
in the polymer products. In a still further aspect, the undesired
branching point can be alleviated through two main methods:
kinetically, as previously described, and through the use of
glycidol derivatives.
[0212] In a further aspect, the use of the glycidol derivatives
forces the epoxide ring to open exclusively into the L.sub.1,3
orientation. While this is undesired in certain glycidol
homopolymer embodiments, the absence of the primary hydroxyl group
does not allow branching of the polymer to take place. In a still
further aspect, the absence of the primary hydroxyl group increases
the linearity of the polymer product and, when coupled with
depressed reaction temperatures, gives polymers with very small
degrees of branching. In a yet further aspect, the use of protected
glycidol units can yield polymers that are completely linear and
can subsequently be deprotected to yield linear glycidol polymers
with the restored primary hydroxyl groups. [29]
[0213] The objective of the following example was to form a polymer
with the increased solubility of glycidol-based polymers but the
biodegradability of polyesters. In a further aspect, a molecule was
formulate that incorporated the rapid reaction rate of glycidol
with the physiological degradability of polyesters. In a still
further aspect, this formulation would allow incorporation of the
desired characteristics without sacrificing the low reaction
conditions needed to impart a high degree of linearity into the
system.
[0214] In a first trial, glycidol was reacted directly with
4-pentenoyl chloride. Despite the use of pyridine, the excess acid
formed in the reaction was enough to cause an opening of the
strained epoxide ring, yielding a mixture of products, none of
which were desired, as represented by the reaction scheme
below.
##STR00137##
[0215] In a subsequent trial, an alternative method was used in
which a diallyl intermediate was employed following the reaction
scheme below.
##STR00138##
[0216] In order to achieve this molecule, 4-pentenoyl chloride was
reacted with 3-buten-1-ol in a 1:1 ratio to yield a diallyl species
with an ester in the center. This molecule was then oxidized using
m-CPBA to afford a clear liquid product that was determined to have
an epoxide on the oxygen side of the ester while maintaining the
allyl functionality on the carbonyl side.
[0217] As shown in FIG. 14, this new species was confirmed through
2-D NMR techniques and appeared poised to overcome both the
degradability problems of homoglycidol systems as well as the
homoglycidol system's lack of post-modification units.
[0218] Next, in order to force linearity into the system, a
protected glycidol derivative was added to the list of possible
monomers. Ethoxyethyl glycidyl ether (EEGE) was chosen as a viable
candidate as its protected side arm is similar in bulk to that of
the newly synthesized glycidyl ester allyl (GEA), following the
reaction scheme below.
##STR00139##
[0219] With the similar steric bulkiness, it was believed that both
EEGE and GEA will polymerize at similar rates, allowing for a
controlled copolymerization of the two monomers. In a further
aspect, the presence of the EEGE can also allow for subsequent
deprotection, which can yield a completely linear polymer with a
plethora of hydroxyls, esters, and allyls.
[0220] Synthesis of Degradable and Non-Degradable Glycidol Based
Copolymers
[0221] To remedy the physiological degradability issues as well as
the post-modification limitations of the glycidol homopolymers, a
range of copolymers was produced. First, the degradability of the
synthesized polymers was increased and subsequently an increased
degree of post-modification units was introduced. These two
problems were first addressed individually and then a more complete
method was devised.
[0222] For increased degradability, first attempts were aimed at
the incorporation of polyester sections into the backbone of the
polymers through the incorporation of 6-valerolactone (VL) as a
comonomer with the glycidol monomer, according to the reaction
scheme below.
##STR00140##
[0223] However, the stringent reaction conditions that yield the
lowest glycidol branching do not allow for a high incorporation of
the lactones. The decision was made to give up some of the control
over the branching in order to increase the lactone incorporation.
Unfortunately, the lowest temperature at which the polymerization
could be run without the lactone freezing was 10.degree. C. Even at
these elevated temperatures, the large difference in polymerization
kinetics did not allow for a high degree of incorporation.
[0224] Next, allyl glycidyl ether (AGE) was used to introduce allyl
functionalities into the backbone of the polymer, thus alleviating
the poor post-modification potential of poly(glycidol). This
reaction, shown in the reaction scheme below, was successful,
yielding polymers with allyl units dispersed throughout the
structure.
##STR00141##
[0225] However the lack of a readily degradable unit meant that the
synthesized polymers could serve little purpose apart from
illuminating the cross-linking ability of the new, semi-branched
structures.
[0226] Next, the newly synthesized GEA monomer was incorporated
into glycidol, according to the reaction scheme below.
##STR00142##
[0227] Unfortunately, the successful synthesis of the novel GEA
monomer species was followed by the monomer's lack-luster
performance when copolymerized with glycidol. The drastic
difference in polymerization kinetics, glycidol being very fast and
GEA being rather slow, afforded a polymer product with truncated
incorporation of the GEA monomer, appearing as a fifth of what was
intended. This realization led to the realization that a kinetic
study of the GEA homopolymer is needed so that the optimal reaction
conditions for the new monomer can be discovered. The NMR of
GLY/GEA polymer is shown in FIG. 15.
[0228] Synthesis of One-Pot Block Copolymer Structures
[0229] After determining that glycidol greatly outcompetes many
lactone comonomers, it was proposed that a glycidol homopolymer
capped with ester and allyl-containing units would be beneficial.
In order to accomplish this, glycidol polymers were formed
according to the determined polymerization restrictions and small
amounts of .alpha.-allyl-.delta.-valerolactone (AVL) were added
during the last hour of the polymerization, as the reaction was
allowed to return to room temperature. This subsequent addition of
the degradable lactone monomer was expected to add onto the end of
the already formed poly(glycidol). The reaction scheme is shown
below.
##STR00143##
[0230] While .sup.1HNMR does show the inclusion of ally groups to
the polymer product, since glycidol is such a kinetically favored
monomer, it cannot be determined if the product is actually a block
copolymer or a random copolymer with a large glycidol "tail."
[0231] Regardless of the actual morphology of the polymer product,
the incorporation of the allyl groups should allow for subsequent
conjugation to free thiols on the exterior of biological structures
such as proteins. It is believed that the attachment of these
degradable hydrophilic polymers to proteins will increase the
proteins' solubility and provide a system that is more advantageous
for protein delivery than the PEGylated protein structures that are
currently in use.
[0232] Synthesis of Linear Polyesters
[0233] Next, formation of linear polyester systems was
investigated. [7-10] In one aspect, it was unknown whether tin
triflate would yield polyester polymers similar to the ones
obtained using the previously employed tin ethyhlhexanoate. In
another aspect, tin triflate is more reactive than tin
ethylhexanoate due to the large electron withdrawing character of
its ligands. Therefore, in a further aspect, tin triflate is a
preferential catalyst if it allows for the same control over
polymer size and PDI as tin ethylhexanoate. First trials were
performed using VL and AVL as copolymers according to the reaction
scheme below.
##STR00144##
[0234] The produced linear polymers exhibited correct size and
distribution with a faster reaction time and the ability to run the
reaction at room temperature rather than elevated temperatures.
Upon this positive outcome, further implementation of tin triflate
was employed.
[0235] As depicted in FIG. 16, the addition of 2-oxepane-1,5-dione
(OPD) to the backbone of the linear polyester systems imparts a
higher degree of water solubility to the system.[7, 9, 10] Since
this increased hydrophilicity is beneficial for the eventual use of
the linear polyesters as the building blocks for nanoparticle drug
delivery systems, a range of OPD containing VL/AVL polymers were
evaluated to study its influence on the system.
[0236] The resulting polymer products contained OPD percentages
ranging from 5%-40%. Furthermore, the purification of the polymer
had to undergo a change. Rather than precipitating in methanol, as
done with VL/AVL polymers, the new OPD containing polymers must be
dialyzed against DCM as they do not precipitate in methanol. As
expected, with the increase in OPD, there was an increase in the
degree of water solubility. These new OPD polymers will be used for
the formation of nanoparticles with increased hydrophilicity that
will be employed as drug delivery vehicles. The exemplary reaction
scheme below shows the synthesis of VL/AVL/OPD linear polymer
according to the above method.
##STR00145##
[0237] Synthesis of Novel Crosslinker Molecules for the Formation
of Nanoparticles
[0238] As described herein, the present invention, in one aspect,
also involves novel cross-linking molecules. In a further aspect,
the cross-linking molecules have the ability to be protonated. In
another aspect, the present invention also relates to the delivery
of biological structures with the nanoparticles using the
cross-linking molecules. In a further aspect, the protonation
capacity is important since biological structures, such as siRNA
will be held more tightly by the positive charges than they would
be if the cross-linking of the polymers was the only method being
employed to contain the biological structures in the nanoparticles.
In a still further aspect, the efficacy of this approach has
recently been illuminated in dendritic polyglycerol
species.[37]
[0239] In a first example, a protected dithiol species was
employed. The disulfide was attached to a carboxylic acid and
subsequently reacted with 0.5 equivalents of a diamine species,
affording a molecule with secondary amine species and readily
accessible thiol groups. The thiol groups were used for "click"
reactions[8] in order to form nanoparticles (FIG. 17), while the
secondary amines were used to increase the complexation of siRNA
into the system (FIG. 18). [37, 40] The exemplary reaction scheme
below shows the formation of a nanoparticle in accordance with the
above method.
##STR00146##
[0240] The final step in the formation of the nanoparticle drug
delivery system is the collapsing of the polymers into
nanoparticles. In one aspect, this reaction was conducted by two
separate methods. In a further aspect, the methods include
thiolene-click reactions[8](FIG. 19) and amine-epoxide reactions[7,
10](FIG. 20). In still further aspect, the two methods can also be
employed in the formation of polyester nanoparticles as well as the
synthesis of novel polyglycerol structures. The exemplary reaction
scheme in FIG. 21 shows thiolene-click GLY/AGE nanoparticle
formation.
[0241] Next, to evaluate the viability of the nanoparticle systems,
drug loading and release studies were performed. As depicted in
FIG. 22, a number of small molecule drugs including Paclitaxel,
Bromonidine, and Temozolomide, were chosen to be loaded into the
polyester nanoparticles. The percentage of drug incorporation was
determined using a nano-drop method based on UV/VIS absorption. The
drug release profiles of these systems was studied by dissolving
the loaded nanoparticles in a physiological pH buffer and allowing
the solution to stir at 37.degree. C. The buffer was changed every
48 hours and the excreted drug was extracted using DCM and
quantified using nanoDrop. Similar techniques will be employed in
order to study the drug loading and release capabilities of the
newly synthesized poly(glycidol) based systems.
[0242] Dual Component Delivery System
[0243] In various aspects, the present invention also relates to a
two component delivery system. In a further aspect, the dual
component drug delivery system can deliver 2 classes of
therapeutics. For example, in one aspect, a dual component drug
delivery system can help achieve bone union following fracture in
patients with neurofibromatosis (NF1). These patients cannot heal
their bone and require amputation. In a further aspect, mouse
models can recapitulate this skeletal complication. Thus, in a
still further aspect, in this example the objective was to combine
small molecules, such as MEK inhibitors, and BMP2 growth factors to
promote bone union (FIG. 27).
[0244] In another aspect, the present invention provides a
reconfigurable and responsive network system (FIG. 28). In a
further aspect, the network systems comprise functionalized
polyglycidol-based crosslinking materials for hydrogels with
functionalized polyesters or polycarbonates. In a still further
aspect, the network systems can comprise functionalized
polyglycidols crosslinked with functionalized degradable materials
such as linear polyesters and polycarbonates. In one aspect, the
networks can be reconfigured and are not thermosets, but rather act
as vitrimers.
These networks are not "set" in the presence of the Zn(Ac).sub.2,
and the free --OH groups of the polyclidol can react with available
esters, making the polymers therefore stimuli responsive. In an
even further aspect, the networks systems are reconfigurable and
are not set. Exemplary reactions for forming functionalized
polyglycidols in a network system are shown in FIG. 29.
I. EXPERIMENTAL EXAMPLES
[0245] The following examples are put forth so as to provide those
of ordinary skill in the art with a complete disclosure and
description of how the compounds, compositions, articles, devices
and/or methods claimed herein are made and evaluated, and are
intended to be purely exemplary of the invention and are not
intended to limit the scope of what the inventors regard as their
invention. Efforts have been made to ensure accuracy with respect
to numbers (e.g., amounts, temperature, etc.), but some errors and
deviations should be accounted for. Unless indicated otherwise,
parts are parts by weight, temperature is in .degree. C. or is at
ambient temperature, and pressure is at or near atmospheric.
[0246] Several methods for preparing the compounds of this
invention are illustrated in the following Examples. Starting
materials and the requisite intermediates are in some cases
commercially available, or can be prepared according to literature
procedures or as illustrated herein.
[0247] The following exemplary compounds of the invention were
synthesized. The Examples are provided herein to illustrate the
invention, and should not be construed as limiting the invention in
any way. The Examples are typically depicted in free base form,
according to the IUPAC naming convention. However, some of the
Examples were obtained or isolated in salt form.
[0248] As indicated, some of the Examples were obtained as racemic
mixtures of one or more enantiomers or diastereomers. The compounds
may be separated by one skilled in the art to isolate individual
enantiomers. Separation can be carried out by the coupling of a
racemic mixture of compounds to an enantiomerically pure compound
to form a diastereomeric mixture, followed by separation of the
individual diastereomers by standard methods, such as fractional
crystallization or chromatography. A racemic or diastereomeric
mixture of the compounds can also be separated directly by
chromatographic methods using chiral stationary phases.
1. General Methods
[0249] All reagent chemicals were purchased from Sigma-Aldrich,
Strem Chemicals, or Acros and used as received unless otherwise
noted. The m-CPBA (77%) was purified as previously reported in the
literature while .delta.-valerolactone and glycidol was further
purified through vacuum distillation. SnakeSkin.RTM. Pleated
Dialysis Tubing, regenerated cellulose, was purchased from Pierce
Biotechnology. Spectra/Por.RTM. Dialysis membrane was purchased
from Spectrum Laboratories Inc.
.alpha.-allyl-.delta.-valerolactone,
.alpha.-propargyl-.delta.-valerolactone, and 2-oxepane-1,5-dione
were synthesized as previously reported in the literature.
[0250] .sup.1H and .sup.13C NMR were obtained from a Bruker AV-I
400 MHz, a Bruker DRX 500 MHz, or a Bruker AV-II 600 MHz
spectrometer. The reported chemical shifts are in ppm and are in
reference to the corresponding residual nuclei in deuterated
solvents.
[0251] Gel-permeation chromatography (GPC) was carried out with a
Waters chromatograph system equipped with a Waters 2414 refractive
index detector, a Waters 2481 dual X absorbance detector, a Waters
1525 binary HPLC pump, and four 5 mm Waters columns (300
mm.times.7.7 mm), connected in series with increasing pore size
(100, 1000, 100,000 and 1,000,000 .ANG. respectively). All runs
were performed with N--N-dimethylformamide (DMF) as the eluent at a
flow rate of 1 mL/min.
[0252] Samples for transmission electron microscopy (TEM) imaging
were prepared by dissolving 0.5 mg nanoparticles in 1 mL
isopropanol, 0.3 mL acetonitrile and 0.2 mL toluene. The samples
were sonicated for 5 min and were stained with 3 drops of 3%
phosphotungstic acid. The carbon grids were prepared by slowly
dipping an Ultrathin Carbon Type-A 400 Mesh Copper Grid (Ted Pella,
Inc., Redding, Calif.) into the particle solutions three times and
drying the grid at ambient temperature. A Philips CM20T
transmission electron microscope operating at 200 kV in
bright-field mode was used to obtain TEM micrographs of the
polymeric nanoparticles. FIG. 23 shows the DLS (Nanosight
instrument), depicting the minimal size dispersity of nanoparticle
structures (115 nm).
2. Preparation of Glycidol Polymers
a. General Procedure
[0253] A 25 mL and a 10 mL round bottom flask were flame-dried
under N.sub.2(g), along with a 50 mL 3-neck round bottom flask
equipped with a stir bar. In the 25 mL round bottom flask, an 1.7M
Iso-Amyl alcohol (IAOH) stock solution was formed using dry THF,
while the 10 mL round bottom flask was used to create a
3.7.times.10.sup.-2M tin triflate stock solution, also using dry
THF. The stock solutions were then allowed to sit for 30 minutes
before adding Sn(OTf).sub.2 (0.00035 eq) and IOAH (0.066 eq) to the
reaction flask. The reaction flask was then brought to the proper
reaction temperature before adding the monomers (1.0 and allowing
the polymerization to run to completion. Figures are shown using an
ethanol initiator (EtOH) instead of the IAOH. The use is optional
but the IAOH is preferred because the polymer products can be more
accurately characterized. The protons from IAOH do not overlap with
polymer peaks and the polymer molecular weight can be determined
via .sup.1H-NMR.
Preparation of GLY Homopolymer
##STR00147##
[0255] Sn(OTf).sub.2 (0.26 mL; 9.45.times.10.sup.-6 mol; 0.00035
eq) and IAOH (0.20 mL; 3.33.times.10.sup.-4 mol; 0.066 eq) were
added to the reaction flask, which was then lowered into an
acetone/dry ice bath at -42.degree. C. The flask was allowed to
cool completely before adding the glycidol (2.00 g; 27.00 mmol; 1.0
eq) monomer. The reaction was then allowed to run for 8 h while
maintaining the depressed temperature. After 8 h the resulting
viscous polymer product was precipitated into vigorously stirring
hexanes affording a clear viscous product. The hexanes were
decanted from the product, which was then transferred to a 6-dram
vial, using methanol. Yield: 1.896 g (94.82%). .sup.1H-NMR (600
MHz, CDCl.sub.3) .delta.: 3.31-3.94 (6H). .sup.13C-NMR (150 MHz,
CDCl.sub.3) .delta.: 81.37, 79.81, 75.12, 73.88, 72.01-72.94,
70.42-71.17, 64.41, 62.53, 62.06.
(1) Preparation of GLY/AGE Polymer (80/20)
##STR00148##
[0257] Sn(OTf).sub.2 (0.23 mL; 8.52.times.10.sup.-6 mol; 0.00035
eq) and EtOH (0.196 mL; 3.33.times.10.sup.-4 mol; 0.066 eq) were
added to the reaction flask, which was then lowered into an
acetonitrile/dry ice bath at -42.degree. C. The flask was allowed
to cool completely before adding the glycidol (1.44 g; 19.47 mmol;
4.0 eq) and glycidyl ether (0.56 g; 4.87 mmol; 1.0 eq). The
reaction was then allowed to run for 8 h while maintaining the
depressed temperature. After 8 h the resulting viscous polymer
product was precipitated into vigorously stirring hexanes affording
a clear viscous product. The hexanes were decanted from the
product, which was then transferred to a 6-dram vial, using
methanol. Yield: 2.05 g (68.27%). .sup.1H-NMR (600 MHz, CDCl.sub.3)
.delta.: 5.92 (1H), 5.21 (2H), 4.04 (2H), 3.38-3.94 (27.30H).
.sup.13C-NMR (150 MHz, CDCl.sub.3) .delta.: 136.31, 117.42, 81.56,
80.01, 74.09, 73.43, 72.51, 70.87, 64.60, 62.69.
(2) Preparation of GEA Homopolymer
##STR00149##
[0259] Sn(OTf).sub.2 (0.056 mL; 2.06.times.10.sup.-6 mol; 0.00035
eq) and IAOH (0.098 mL; 1.67.times.10.sup.-4 mol; 0.066 eq) were
added to the reaction flask, which was then lowered into an
acetonitrile/dry ice bath at -42.degree. C. The flask was allowed
to cool completely before adding the previously synthesized
glycidyl ester allyl (1.0 g; 5.88 mmol; 1.0 eq). The reaction was
then allowed to run for 8 h while maintaining the depressed
temperature. After 8 h the resulting viscous polymer product was
precipitated into vigorously stirring hexanes affording a clear
viscous product. The hexanes were decanted from the product, which
was then transferred to a 6-dram vial, using methanol. Yield: 230.8
mg (23.08%). .sup.1H-NMR (600 MHz, CDCl.sub.3) .delta.: 5.83 (1H),
5.00 (2H) 4.61 (5H), 3.39-3.82 (19.06H), 2.24-2.66 (23.33H) 2.12
(6.17H) 1.56-1.92 (4.87H). .sup.13C-NMR (150 MHz, CDCl.sub.3)
.delta.: 180.40, 174.87, 138.14, 136.53, 135.64, 117.62, 115.98,
81.31, 79.58, 70.84, 67.32, 66.52, 64.62, 60.05, 37.16, 34.39,
31.13, 29.55, 24.51.
(3) Preparation of GLY/GEA Polymer (80/20)
##STR00150##
[0261] Sn(OTf).sub.2 (0.10 g; 3.75.times.10.sup.-6 mol; 0.00035 eq)
and EtOH (0.10 mL; 1.67.times.10.sup.-4 mol; 0.066 eq) were added
to the reaction flask, which was then lowered into an
acetonitrile/dry ice bath at -42.degree. C. The flask was allowed
to cool completely before adding the glycidol (0.64 g; 8.57 mmol;
4.0 eq) and the previously synthesized glycidyl ester allyl (0.36
g; 2.14 mmol; 1.0 eq). The reaction was then allowed to run for 8 h
while maintaining the depressed temperature. After 8 h the
resulting viscous polymer product was precipitated into vigorously
stirring hexanes affording a clear viscous product. The hexanes
were decanted from the product, which was then transferred to a
6-dram vial, using methanol. Yield: 633.7 mg (63.37%). .sup.1H-NMR
(600 MHz, CDCl.sub.3) .delta.: 5.93 (1H), 5.27 (2.37H), 4.06-4.28
(2.91H) 3.26-3.98 (303.6H), 2.24-2.63 (8.25H), 1.84 (2.57H), 1.65
(18.67H). .sup.13C-NMR (150 MHz, CDCl.sub.3) .delta.: 138.53,
72.90, 71.32, 63.42, 61.52, 26.55.
(4) Preparation of GLY/MLGEA Polymer (80/20)
##STR00151##
[0263] Sn(OTf).sub.2 (0.10 g; 3.75.times.10.sup.-6 mol; 0.00035 eq)
and EtOH (0.10 mL; 1.67.times.10.sup.-4 mol; 0.066 eq) were added
to the reaction flask, which was then lowered into an
acetonitrile/dry ice bath at -42.degree. C. The flask was allowed
to cool completely before adding the glycidol (0.64 g; 8.84 mmol;
4.0 eq) and the previously synthesized mixed-length glycidyl ester
allyl (0.35 g; 2.21 mmol; 1.0 eq). The reaction was then allowed to
run for 8 h while maintaining the depressed temperature. After 8 h
the resulting viscous polymer product was precipitated into
vigorously stirring hexanes affording a clear viscous product. The
hexanes were decanted from the product, which was then transferred
to a 6-dram vial, using methanol. .sup.1H-NMR (600 MHz, CDCl.sub.3)
.delta.: 5.83 (1H), 5.07 (2.37H), 3.97-4.21 (1.74H) 3.29-3.95
(97.12H), 2.26-2.61 (4.95H), 1.82 (1.61H), 1.65 (7.54H).
(5) Preparation of EEGE Homopolymer
##STR00152##
[0265] Sn(OTf).sub.2 (0.032 mL; 1.20.times.10.sup.-6 mmol; 0.00035
eq) and EtOH (0.049 mL; 8.33.times.10.sup.-5 mmol; 0.066 eq) were
added to the reaction flask. The synthesized ethoxyethyl glycidyl
ether (0.5 g; 3.42 mmol; 1.0 eq) was then added. The reaction was
then allowed to run for 24 h at room temperature. After 24 h the
resulting viscous polymer product was precipitated into vigorously
stirring hexanes affording a whitish viscous product. Yield: 196.5
mg (39.3%). The hexanes were decanted from the product, which was
then transferred to a 6-dram vial, using methanol. .sup.1H-NMR (500
MHz, CDCl.sub.3) .delta.: 4.16 (1H), 3.87 (1.21H), 3.39-3.81
(19.28H), 1.64 (5.54H), 1.23 (8.08H), 0.86 (2.92H).
(6) Preparation of EEGE/GEA Polymer (80/20)
##STR00153##
[0267] Sn(OTf).sub.2 (0.0313 mL; 1.16.times.10.sup.-6 mmol; 0.00035
eq) and EtOH (0.049 mL; 8.33.times.10.sup.-5 mmol; 0.066 eq) were
added to the reaction flask. The stock solutions were allowed to
stir for 10 minutes before adding the .delta.-valerolactone (1.48
g; 14.80 mmol; 4.0 eq) and .alpha.-allyl-.delta.-valerolactone
(0.52 g; 3.70 mmol; 1.0 eq). The reaction was then allowed to run
for 24 h at room temperature. After 24 h the resulting viscous
polymer product was precipitated into cold diethyl ether to afford
the off-white particulate polymer product. The diethyl ether was
decanted from the product, which was then transferred to a 6-dram
vial, using ethyl acetate. Yield: 374.9 mg (74.98%). .sup.1H-NMR
(500 MHz, CDCl.sub.3) .delta.: 4.28 (1H), 3.43-3.78 (6.43H), 3.17
(1.32H), 2.35 (3.36H), 2.02 (2.84H), 1.65 (10.42H), 1.29 (69.79H),
0.92 (37.31H).
(7) Preparation of EEGE/MLGEA Polymer (80/20)
##STR00154##
[0269] Sn(OTf).sub.2 (0.0313 mL; 1.16.times.10.sup.-6 mmol; 0.00035
eq) and EtOH (0.049 mL; 8.33.times.10.sup.-5 mmol; 0.066 eq) were
added to the reaction flask. The stock solutions were allowed to
stir for 30 minutes before adding the EEGE (1.36 g; 14.54 mmol; 4.0
eq) and MLGEA (0.47 g; 3.36 mmol; 1.0 eq). The reaction was then
allowed to run for 24 h at 70.degree. C. After 24 h the resulting
viscous polymer product was precipitated into methanol to afford
the off-white particulate polymer product. The methanol was
decanted from the product, which was then transferred to a 6-dram
vial, using ethyl acetate.
(8) Preparation of GLY/VL Polymer (80/20)
##STR00155##
[0271] Sn(OTf).sub.2 (024 mL; 8.82.times.10.sup.-6 mmol; 0.00035
eq) and EtOH (0.20 mL; 3.33.times.10.sup.-4 mmol; 0.066 eq) were
added to the reaction flask, which was then lowered into a salt
water/ice bath at -20.degree. C. The flask was allowed to cool
completely before adding the glycidol (1.49 g; 20.16 mmol; 4.0 eq)
and .delta.-valerolactone (0.50 g; 5.04 mmol; 1.0 eq). The reaction
was then allowed to run for 8 h while maintaining the depressed
temperature. After 8 h the resulting viscous polymer product was
precipitated into vigorously stirring hexanes affording a whitish
viscous product. The hexanes were decanted from the product, which
was then transferred to a 6-dram vial, using methanol. Yield: 1.54
g (77%). .sup.1H-NMR (600 MHz, CDCl.sub.3) .delta.: 4.10 (1H), 3.89
(1.17H), 3.41-3.82 (18.48H), 3.33 (7.66H), 2.37 (1.36H, 1.51-1.76
(4.29H). .sup.13C-NMR (150 MHz, CDCl.sub.3) .delta.: 138.52, 72.96,
71.38, 69.70, 64.07, 33.50, 28.15, 21.59.
(9) Preparation of GLY/VL/AVL Polymer (60/20/20)
##STR00156##
[0273] Sn(OTf).sub.2 (0.20 mL; 7.56.times.10.sup.-6 mmol; 0.00035
eq) and EtOH (0.20 mL; 3.33.times.10.sup.-4 mmol; 0.066 eq) were
added to the reaction flask, which was then lowered into a salt
water/ice bath at -20.degree. C. The flask was allowed to cool
completely before adding the glycidol (0.96 g; 12.96 mmol; 3.0 eq),
.delta.-valerolactone (0.61 g; 4.32 mmol; 1.0 eq), and
.alpha.-allyl-.delta.-valerolactone (0.43 g; 4.32 mmol; 1.0 eq).
The reaction was then allowed to run for 8 h while maintaining the
depressed temperature. After 8 h the resulting viscous polymer
product was precipitated into vigorously stirring hexanes affording
a whitish viscous product. The hexanes were decanted from the
product, which was then transferred to a 6-dram vial, using
methanol. Yield: 528.4 mg (35.23%). .sup.1H-NMR (600 MHz,
CDCl.sub.3) .delta.: 5.77 (1H), 5.05 (2H), 4.13 (7.46H), 3.29-4.03
(171.73H), 2.38 (14.37H), 1.66 (34.73H), 1.22 (7.25H). .sup.13C-NMR
(150 MHz, CDCl.sub.3) .delta.: 175.82, 136.74, 117.21, 83.16,
81.40, 79.83, 73.85, 72.25, 64.44, 62.45, 37.65, 34.53, 32.94,
31.29, 30.04, 29.24, 27.44, 22.46, 15.49, 14.63.
(10) Preparation of GLY/VL/PO/AGE Polymer (50/20/20/10)
##STR00157##
[0275] Sn(OTf).sub.2 (5.83 mg; 1.28.times.10.sup.-5 mol; 0.00035
eq) and EtOH (0.012 mL; 2.07.times.10.sup.-4 mol; 0.066 eq) were
added to the reaction flask, which was then lowered into a salt
water/ice bath at -20.degree. C. The flask was allowed to cool
completely before adding the glycidol (0.47 g; 6.38 mmol; 5.0 eq),
.delta.-valerolactone (0.26 g; 2.56 mmol; 2.0 eq), propylene oxide
(0.15 g; 2.56 mmol; 2.0 eq), and allyl glycidyl ether (0.15 g; 1.28
mmol; 1.0 eq). The reaction was then allowed to run for 8 h while
maintaining the depressed temperature. After 8 h the resulting
viscous polymer product was precipitated into vigorously stirring
hexanes affording a whitish viscous product. The hexanes were
decanted from the product, which was then transferred to a 6-dram
vial, using methanol. Yield: 218.2 mg (21.82%). .sup.1H-NMR (600
MHz, CDCl.sub.3) .delta.: 5.92 (1H), 5.22 (2.05H), 4.03 (2.36H),
3.29-3.96 (54.76H), 2.16 (1.09H), 1.68 (1.81H), 1.30 (3.74H), 1.15
(10.64H). .sup.13C-NMR (150 MHz, CDCl.sub.3) .delta.: 207.26,
176.02, 136.26, 117.50, 7.70, 74.01, 73.39, 72.28, 70.73, 62.51,
52.22, 34.59, 33.02, 22.53.
(11) Oxidation of GLY/AGE Polymer (80/20)
##STR00158##
[0277] To a round bottom flask equipped with a stir bar, was added
the GLY/AGE polymer (1.0 eq), m-CPBA (0.1 eq), and methanol
(5.4.times.10.sup.-2 g/mL). The round bottom flask was then capped
with a septum and allowed to stir for 72 h at room temperature. The
resulting product solution was then concentrated and precipitated
into vigorously stirring hexanes. The hexane was decanted from the
product, which was then transferred to a 6-dram vial using
methanol. Yield: 128.2 mg (24.97%). .sup.1H-NMR (600 MHz,
CDCl.sub.3) .delta.: 5.93 (1H), 5.26 (2.01H), 4.03 (1.96H), 3.89
(4.00H), 3.38-3.81 (53.76H), 3.31 (1.07H), 1.65 (5.33H).
.sup.13C-NMR (150 MHz, CDCl.sub.3) .delta.: 136.26, 117.40, 81.47,
79.92, 79.27, 74.02, 72.41, 70.72, 64.54, 62.63, 27.73.
3. Preparation of Monomers
(1) Purification of m-CPBA
[0278] m-CPBA (70 g; 77%) was dissolved in diethyl ether (500 mL)
and transferred to a separatory funnel. The ether layer was then
washed 3.times. with 300 mL aliquots of buffer solution (410 mL
0.1M NaOH, 250 mL 0.2M KH.sub.2PO.sub.4, made up to 1 L;
pH.apprxeq.7.5). The ether layer was dried over MgSO.sub.4 and then
evaporated on the rotovap to yield the pure white m-CPBA
product.[41].sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 8.14-8.08
(2H, m, CH, CH), 7.82 (1H, d, CH), 7.59 (1H, m, CH).
(2) Preparation of a-Allyl-.DELTA.-Valerolactone
##STR00159##
[0280] This reaction was performed as previously described in
literature with the added use of vacuum distilled
.delta.-valerolactone rather than using the purchased purity.[7, 9,
10] Yield: 2.56 g (46.12%).sup.1H-NMR (400 MHz, CDCl.sub.3)
.delta.: 5.80 (1H, m, CH) 5.04 (2H, m, CH.sub.2), 4.27 (2H, m,
CH.sub.2), 2.42 (2H, m, CH.sub.2, CH), 2.18 (1H, m, CH.sub.2),
2.00-1.72 (4H, m, CH.sub.2). .sup.13C-NMR (100 MHz, CDCl.sub.3)
.delta.: 173.98, 135.03, 117.54, 68.84, 39.13, 35.42, 22.13,
21.05.
(3) Preparation of a-Propargyl-.DELTA.-Valerolactone
##STR00160##
[0282] This reaction was performed as previously described in
literature with the added use of vacuum distilled
.delta.-valerolactone rather than using the purchased purity.[7, 9,
10] Yield: 2.08 g (53.64%). .sup.1H-NMR (400 MHz, CDCl.sub.3)
.delta.: 4.34 (2H, m, CH.sub.2), 2.71 (1H, m, CH), 2.51 (1H, m,
CH.sub.2), 2.37 (1H, m, CH), 2.24 (1H, m, CH.sub.2), 2.00-1.73 (4H,
m, CH.sub.2). .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta.: 171.96,
81.75, 70.14, 68.98, 39.02, 24.21, 21.87, 20.43.
(4) Preparation of 2-Oxepane-1,5-Dione
##STR00161##
[0284] This reaction was performed as previously described in
literature with the added use of purified m-CPBA rather than using
the purchased purity.[7, 9, 10] Yield: 1.89 g (54.31%). .sup.1H-NMR
(400 MHz, CDCl.sub.3) .delta.: 4.36 (2H, t, CH.sub.2), 2.76 (2H, m,
CH.sub.2), 2.65 (2H, m, CH.sub.2), 2.59 (2H, m, CH.sub.2).
.sup.13C-NMR (125 MHz, CDCl.sub.3) .delta.: 205.14, 173.57, 63.74,
44.86, 38.94, 28.31.
(5) Preparation of Diallyl Ester
##STR00162##
[0286] To a round bottom flask equipped with a stir bar, was added
3-buten-1-ol (3.27 g; 45.29 mmol; 1.0 eq) and DCM (25 mL; excess).
A diluted solution of 4-pentenoyl chloride (5.37 g; 45.29 mmol; 1.0
eq) and DCM (25 mL; excess) was created in an addition funnel. The
4-pentenoyl chloride solution was then added drop wise to the
stirring reaction mixture over 30 minutes and the reaction was
allowed to run for 3 h until TLC indicated the reaction was
complete. The excess solvent was then removed on the rotovap to
afford the crude product. The resulting crude liquid product was on
the Biotage column system using a gradient of 8%-70% ethyl acetate
in hexanes to yield the pure clear liquid product. Yield: 10.33 g
(73.95%). .sup.1H-NMR (500 MHz, CDCl.sub.3/TMS) .delta. 5.78 (2H,
m, CH), 5.07 (4H, m, CH.sub.2), 4.12 (2H, t, CH.sub.2O), 2.38 (6H,
m, 3CH.sub.2). .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta. 173.22,
136.89, 134.22, 117.38, 115.65, 63.61, 33.73, 33.28, 29.07.
(6) Preparation of Glycidyl Ester Allyl
##STR00163##
[0288] To a round bottom flask equipped with a stir bar, was added
the previously synthesized diallyl ester (2.90 g; 18.79 mmol; 1.0
eq), m-CPBA (3.24 g; 18.79 mmol; 1.0 eq), and DCM (53.66 mL;
5.4.times.10.sup.-2 g/mL). The oxidation reaction was then allowed
to run for 48 h. The crude product was then vacuum filtered to
remove the white precipitate before extracting the filtrate with
saturated sodium bicarbonate to remove any unreacted m-CPBA. The
excess DCM was then removed on the rotovap to afford the clear
crude liquid product. The resulting crude liquid product was
purified on the Biotage column system using a gradient of 8%-70%
ethyl acetate in hexanes to yield the pure clear liquid product.
Yield: 6.83 g (60.47%). .sup.1H-NMR (500 MHz, CDCl.sub.3) .delta.
5.82 (1H, m, CH), 5.09 (2H, m, CH.sub.2), 4.41 (1H, dd, CH.sub.2),
3.93 (1H, q, CH.sub.2), 3.21 (1H, sext, CH), 2.85 (1H, t,
CH.sub.2), 2.65 (1H, q, CH.sub.2), 2.47 (2H, m, CH.sub.2), 2.40
(2H, m, CH.sub.2). .sup.13C-NMR (125 MHz, CDCl.sub.3) .delta.
174.27, 138, 116.06, 66.23, 50.42, 45.13, 34.27, 29.96.
(7) Preparation of Mixed Length Diallyl Ester
##STR00164##
[0290] To a round bottom flask equipped with a stir bar, was added
allyl alcohol (3.27 g; 45.29 mmol; 1.0 eq) and DCM (25 mL; excess).
A diluted solution of 4-pentenoyl chloride (5.37 g; 45.29 mmol; 1.0
eq) and DCM (25 mL; excess) was created in an addition funnel. The
4-pentenoyl chloride solution was then added drop wise to the
stirring reaction mixture over 30 minutes and the reaction was
allowed to run for 3 h until TLC indicated the reaction was
complete. The excess solvent was then removed on the rotovap to
afford the crude product. The resulting crude liquid product was
purified on the Biotage column system using a gradient of 8%-70%
ethyl acetate in hexanes to yield the pure clear liquid product.
.sup.1H-NMR (500 MHz, CDCl.sub.3/TMS) .delta. 5.78 (2H, m, CH),
5.09 (4.11H, m, CH.sub.2), 4.12 (2H, t, CH.sub.2O), 2.39 (6.4H, m,
3CH.sub.2).
(8) Preparation of Mixed Length Glycidyl Ester Allyl
##STR00165##
[0292] To a round bottom flask equipped with a stir bar, was added
the previously synthesized mixed length diallyl ester (2.90 g;
18.79 mmol; 1.0 eq), m-CPBA (3.24 g; 18.789 mmol; 1.0 eq), and DCM
(53.657 mL; 5.4.times.10.sup.-2 g/mL). The oxidation reaction was
then allowed to run for 48 h. The crude product was then vacuum
filtered to remove the white precipitate before extracting the
filtrate with saturated sodium bicarbonate to remove any unreacted
m-CPBA. The excess DCM was then removed on the rotovap to afford
the clear crude liquid product. The resulting crude liquid product
was purified on the Biotage column system using a gradient of
8%-70% ethyl acetate in hexanes to yield the pure clear liquid
product. .sup.1H-NMR (500 MHz, CDCl.sub.3) .delta. 5.88 (2H, m,
CH), 5.22 (4.54H, m, CH.sub.2), 2.95 (2.19H, m), 2.72 (2.58H, t),
1.94 (3.06H, m), 1.75, 2.87H).
(9) Preparation of Ethoxyethyl Glycidol Ether
##STR00166##
[0294] To a round bottom flask equipped with a stir bar was added
glycidol (7.41 g; 100 mmol; 1.0 eq) and ethyl vinyl ether (27.88 g;
386.67 mmol; 3.87 eq). The reaction flask was then lowered into a
salt water/ice bath at 0.degree. C. and began stirring. P-toluene
sulfonic acid (185.2 mg; 0.97 mmol; 0.0097 eq) was then added
slowly portionwise in order to maintain the low reaction
temperature. The mixture was then allowed to stir at the depressed
temperature for 7 h. The reaction was then quenched with saturated
NaHCO.sub.3 (excess). The organic layer was then separated, dried
and evaporated on the rotovap to yield to monomer product. Yield:
182.6 mg (32.47%). .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 4.73
(1.95H), 4.09 (1H), 3.78 (1H), 3.66 (3.15H), 3.51 (2.95H), 2.77
(2.01H), 2.61 (1.98H), 1.29 (6.18H), 1.17 (7.97H). .sup.13C-NMR
(100 MHz, CDCl.sub.3) .delta.: 171.21, 99.76, 65.86, 65.19, 60.98,
50.89, 44.64, 21.12, 19.72, 15.35, 14.29.
4. Preparation of Polyesters
a. General Procedure for VL Based Polymers
[0295] A 25 mL and a 10 mL round bottom flask were flame-dried
under N.sub.2(g), along with a 50 mL 3-neck round bottom flask
equipped with a stir bar. In the 25 mL round bottom flask, an 1.7M
EtOH stock solution was formed using dry THF, while the 10 mL round
bottom flask was used to create a 3.7.times.10.sup.-2M tin triflate
stock solution, also using dry THF. The stock solutions were then
allowed to sit for 30 minutes before adding Sn(OTf).sub.2 (0.00035
eq) and EtOH (0.066 eq) to the reaction flask. The reaction flask
was then brought to the proper reaction temperature before adding
the monomers and allowing the polymerization to run to
completion.
(1) Preparation of VL Homopolymer
##STR00167##
[0297] Sn(OTf).sub.2 (0.19 mL; 6.99.times.10.sup.-6 mmol; 0.00035
eq) and EtOH (0.29 mL; 5.times.10.sup.-4 mmol; 0.066 eq) were added
to the reaction flask. The stock solutions were allowed to stir for
10 minutes before adding the .delta.-valerolactone (2.70 g; 19.98
mmol; 1.0 eq) monomer. The reaction was then allowed to run for 24
h at room temperature. After 24 h the resulting viscous polymer
product was precipitated into cold diethyl ether to afford the
off-white particulate polymer product. The diethyl ether was
decanted from the product, which was then transferred to a 6-dram
vial, using ethyl acetate. Yield: 1.83 g (91.5%). .sup.1H-NMR (400
MHz, CDCl.sub.3) .delta.: 4.07 (82.79H), 3.63 (6.44H), 3.46
(101.51H), 2.33 (93.10H), 2.21 (33.20H), 1.67 (168.64H), 1.25 (3H).
.sup.13C-NMR (100 MHz, CDCl.sub.3) .delta.: 173.62, 63.89, 62.18,
50.60, 33.62, 32.06, 27.98, 21.34.
(2) Preparation of VL/AVL Linear Polymer (80/20)
##STR00168##
[0299] Sn(OTf).sub.2 (0.19 mL; 6.99.times.10.sup.-6 mmol; 0.00035
eq) and EtOH (0.29 mL; 5.times.10.sup.-4 mmol; 0.066 eq) were added
to the reaction flask. The stock solutions were allowed to stir for
10 minutes before adding the .delta.-valerolactone (1.48 g; 14.80
mmol; 4.0 eq) and .alpha.-allyl-.delta.-valerolactone (0.52 g; 3.70
mmol; 1.0 eq). The reaction was then allowed to run for 24 h at
room temperature. After 24 h the resulting viscous polymer product
was precipitated into cold diethyl ether to afford the off-white
particulate polymer product. The diethyl ether was decanted from
the product, which was then transferred to a 6-dram vial, using
ethyl acetate. Yield: 1.3815 g (69.08%). .sup.1H-NMR (400 MHz,
CDCl.sub.3) .delta.: 5.72 (1H), 5.03 (2H), 4.08 (11.52H), 2.35
(11.77H), 1.67 (24.01H), 1.27 (1.85H). .sup.13C-NMR (100 MHz,
CDCl.sub.3) .delta.: 175.30, 173.42, 135.36, 117.11, 64.07, 62.37,
60.50, 44.98, 36.59, 33.93, 32.23, 28.23, 26.57, 21.57, 21.28,
14.39.
(3) Preparation of VL/OPD/AVL Linear Polymer (60/20/20)
##STR00169##
[0301] Sn(OTf).sub.2 (0.30 mL; 8.83.times.10.sup.-6 mmol; 0.00035
eq) and EtOH (0.29 mL; 3.33.times.10.sup.-4 mmol; 0.066 eq) were
added to the reaction flask. The stock solutions were allowed to
stir for 10 minutes before adding the .delta.-valerolactone (1.92
g; 19.20 mmol; 3.0 eq), 2-oxepane-1,5-dione (0.82 g; 6.40 mmol; 1.0
eq), and the previously synthesized
.alpha.-allyl-.delta.-valerolactone (0.90 g; 6.40 mmol; 1.0 eq).
The reaction was then allowed to run for 24 h at room temperature.
After 24 h the resulting viscous polymer product was precipitated
into cold diethyl ether to afford the off-white particulate polymer
product. The diethyl ether was decanted from the product, which was
then transferred to a 6-dram vial, using ethyl acetate. Yield:
2.912 g (97.07%). .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 5.71
(1H, m, CH), 5.03 (2H, m, CH.sub.2), 4.34 (2H), 4.08 (10.79H), 3.67
(1.04H), 2.52-2.86 (6.68H), 2.15-2.51 (13.22H), 1.67 (22.63H).
.sup.13C-NMR (100 MHz, CDCl.sub.3) .delta.: 135. 33, 117.73, 69.64,
68.76, 64.11, 39.57, 35.71, 33.89, 30.05, 28.26, 24.36, 22.53,
21.61, 19.33.
(4) Preparation of VL/PVL/OPD Linear Polymer (70/20/10)
##STR00170##
[0303] Sn(OTf).sub.2 (0.26 mL; 9.50.times.10.sup.-6 mmol; 0.00035
eq) and EtOH (0.29 mL; 5.times.10.sup.-4 mmol; 0.066 eq) were added
to the reaction flask. The stock solutions were allowed to stir for
10 minutes before adding the .delta.-valerolactone (1.90 g; 19.00;
7.0 eq), the previously synthesized
.alpha.-propargyl-.delta.-valerolactone (0.75 g; 5.43 mmol; 2.0
eq), and the previously synthesized
.alpha.-propargyl-.delta.-valerolactone 2-oxepane-1,5-dione (0.35
g; 2.71 mmol; 1.0 eq). The reaction was then allowed to run for 24
h at room temperature. After 24 h the resulting viscous polymer
product was precipitated into cold diethyl ether to afford the
off-white particulate polymer product. The diethyl ether was
decanted from the product, which was then transferred to a 6-dram
vial, using ethyl acetate. Yield: 1.58 g (52.58%). .sup.1H-NMR (400
MHz, CDCl.sub.3) .delta.: 4.34 (1.91H), 4.07 (32.58H), 3.67
(1.76H), 3.41 (1.32H), 2.24-2.83 (42.88H), 2.02 (3.03H), 1.67
(63.84H), 1.25 (3H). .sup.13C-NMR (100 MHz, CDCl.sub.3) .delta.:
173.42, 70.36, 64.06, 60.54, 53.59, 44.11, 42.98, 41.58, 33.83,
28.21, 25.25, 21.56, 14.33.
(5) Oxidation of VL/AVL Linear Polymer (80/20)
##STR00171##
[0305] To a round bottom flask equipped with a stir bar, was added
the VL/AVL polymer (2.55 g; 3.87 mmol; 2.0 eq), m-CPBA (0.44 g;
2.15 mmol; 1.0 eq), and dichloromethane (47.2 mL;
5.4.times.10.sup.-2 g/mL). The round bottom flask was then capped
with a septum and allowed to stir for 72 h at room temperature. The
resulting product solution was then concentrated and precipitated
into cold methanol. The methanol was decanted from the product,
which was then transferred to a 6-dram vial using dichloromethane.
Yield: 835.6 mg (64.28%). .sup.1H-NMR (400 MHz, CDCl.sub.3)
.delta.: 5.72 (1H), 5.04 (2.07H), 4.08 (41.05H), 3.66 (2.77H), 2.93
(0.84H), 2.75 (0.95H), 2.29-2.53 (42.04), 1.69 (92.19H), 1.26
(2.15H). .sup.13C-NMR (100 MHz, CDCl.sub.3) .delta.: 173.47,
135.26, 117.16, 64.97, 62.35, 60.49, 44.97, 36.58, 33.83, 32.20,
28.19, 26.56, 21.58, 21.27, 14.38.
(6) Oxidation of VL/OPD/AVL Linear Polymer (60/20/20)
##STR00172##
[0307] To a round bottom flask equipped with a stir bar, was added
the VL/OPD/AVL polymer (0.35 g; 0.52 mmol; 2.0 eq), m-CPBA (51.89
mg; 0.26 mmol; 1.0 eq), and dichloromethane (6.511 mL;
5.4.times.10.sup.-2 g/mL). The round bottom flask was then capped
with a septum and allowed to stir for 72 h at room temperature. The
resulting product solution was then concentrated and precipitated
into cold methanol. The methanol was decanted from the product,
which was then transferred to a 6-dram vial using dichloromethane.
Yield: 367 mg (84.95%). .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.:
5.73 (1H), 5.05 (2.15H), 4.36 (4.29H), 4.11 (14.01H), 3.90 (2.04H),
3.67 (5.04H), 3.43 (1.13H), 2.52-2.86 (21.18H), 2.16-2.52 (18.28H),
1.69 (34.37H), 1.27 (1.59H). .sup.13C-NMR (150 MHz, CDCl.sub.3)
.delta.: 169.68, 134.83, 133.87, 131.26, 129.99, 128.42, 64.12,
53.60, 33.88, 28.24, 21.60.
5. Preparation of New Crosslinkers
(1) Preparation of Disulfide Linker
##STR00173##
[0309] To a round bottom flask equipped with a stir bar, was added
aldrithiol-2 (10.00 g; 1.5 eq), 3-mercaptopropionic acid (3.212 g;
1.0 eq), and MeOH (excess). The yellow reaction was then allowed to
stir for 72 h. The resulting yellow solution was concentrated and
the yellow product was resuspended in dichloromethane and dried
onto silica gel. The product was then purified through column
chromatography using a gradient of 10%-30% ethyl acetate in hexanes
to yield the pure, slightly off-white solid product. Yield: 2.23 g
(34.17%). .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 8.39 (1H),
7.81 (2H), 7.22 (1H), 3.03 (2H), 2.71 (2H). .sup.13C-NMR (100 MHz,
CDCl.sub.3) .delta.: 175.56, 159.39, 149.60, 137.64, 121.46,
120.85, 34.37, 34.24.
(2) Preparation of Disulfide Amine Crosslinker
##STR00174##
[0311] To a round bottom flask equipped with a stir bar, in an ice
bath, was added the previously synthesized disulfide linker (0.50
g; 2.32 mmol; 6.0 eq), THF (excess), and triethylamine (0.33 g;
3.25 mmol; 8.4 eq), followed by the slow addition of isobutyl
chloroformate (0.40 g; 2.90 mmol; 7.5 eq). The reaction was then
allowed to stir for 3 hours. To the stirring reaction mixture was
then added pentaethylenhexamine (0.09 g; 0.39 mmol; 1.0 eq) slowly
drop wise. The reaction was then removed from the ice bath and
allowed to run for an additional 24 hours at room temperature. The
excess solvent was then removed on the rotovap to afford the crude,
deep red product. The resulting crude product was purified on the
Biotage column system using a gradient of 2%-20% ethyl acetate in
hexanes to yield the pure, slightly off-white solid product.
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 8.42 (2H), 7.48 (2H),
7.18 (2.01H), 6.98 (2.15H) 3.90 (4.56H), 3.45 (4.17H), 3.34
(1.16H), 2.89 (1.06H), 2.80 (4.35), 1.13 (0.96H), 0.94 (15.11H).
.sup.13C-NMR (100 MHz, CDCl.sub.3) .delta.: 172.33, 158.41, 149.57,
136.17, 122.62, 119.65, 53.61, 42.93, 34.93, 27.88, 25.18,
19.27.
6. Preparation of Nanoparticles
(1) Nanoparticle Formation Through Thiolene-Click Reactions Using
VL/OPD/AVL Copolymers (50 nm)
[0312] To a round bottom flask equipped with a stir bar, was added
the VL/OPD/AVL polymer (111.5 mg; 5.431.times.10.sup.-2 mmol; 1
eq), 3,6-dioxa-1,8-octanedithiol (9.90 mg; 5.431.times.10.sup.-2
mmol; 1 eq), and dichloromethane (16.76 mL; 3.24.times.10.sup.-3M).
The flask was then fitted with a reflux condenser and lowered into
an oil bath at 45.degree. C. to reflux for 12 h. The resulting
solution was then transferred to 10K dialysis tubing and dialyzed
for 72 h against dichloromethane to remove any unreacted starting
material. The remaining product solution was then concentrated into
a preweighed vial and stored in the fridge. Yield: 81.8 mg
(66.17%). .sup.1H-NMR (600 MHz, CDCl.sub.3) .delta.: The
significant change is the reduction of the allyl peaks at 5.72 and
5.04 ppm and the appearance of signals at 3.65 and 2.69 ppm
corresponding to the protons neighboring the thiols of the PEG
linker after cross-linking All other aspects of the spectrum remain
similar to the polymer spectrum.
(2) Nanoparticle Formation Through Thiolene-Click Reactions Using
GLY/VL/AVL Copolymers (50 nm)
[0313] To a round bottom flask equipped with a stir bar, was added
the GLY/VL/AVL polymer (118.75 mg; 2.74.times.10.sup.-2 mmol; 1
eq), 3,6-dioxa-1,8-octanedithiol (5.00 mg; 2.74.times.10.sup.-2
mmol; 1 eq), and methanol (8.47 mL; 3.24.times.10.sup.-3M). The
flask was then fitted with a reflux condenser and lowered into an
oil bath at 45.degree. C. to reflux for 12 h. The resulting
solution was then transferred to 10K dialysis tubing and dialyzed
for 72 h against dichloromethane to remove any unreacted starting
material. The remaining product solution was then concentrated into
a preweighed vial and stored in the fridge. Yield: 115.6 mg
(93.41%). .sup.1H-NMR (600 MHz, CDCl.sub.3) .delta.: The
significant change is the reduction of the allyl peaks at 5.72 and
5.04 ppm and the appearance of signals at 3.65 and 2.69 ppm
corresponding to the protons neighboring the thiols of the PEG
linker after cross-linking All other aspects of the spectrum remain
similar to the polymer spectrum.
(3) Nanoparticle Formation Through Thiolene-Click Reactions Using
GLY/AGE Copolymers (50 nm)
[0314] To a round bottom flask equipped with a stir bar, was added
the GLY/AGE polymer (111 mg; 1.79.times.10.sup.-1 mmol; 2 eq),
3,6-dioxa-1,8-octanedithiol (16.35 mg; 8.97.times.10.sup.-2 mmol; 1
eq), and methanol (55.36 mL; 3.24.times.10.sup.-3M). The flask was
then fitted with a reflux condenser and lowered into an oil bath at
45.degree. C. to reflux for 12 h. The resulting solution was then
transferred to 10K dialysis tubing and dialyzed for 72 h against
methanol to remove any unreacted starting material. The remaining
product solution was then concentrated into a preweighed vial and
stored in the fridge. Yield: 103.1 mg (84.14%). .sup.1H-NMR (600
MHz, CDCl.sub.3) .delta.: The significant change is the reduction
of the allyl peaks at 5.72 and 5.04 ppm and the appearance of
signals at 3.65 and 2.69 ppm corresponding to the protons
neighboring the thiols of the PEG linker after cross-linking All
other aspects of the spectrum remain similar to the polymer
spectrum. FIG. 24 shows transmission electron microscopy (TEM)
image for GLY/AGE nanoparticles with 3.5% crosslinking, after
running reaction for 12 hours in methanol at 45.degree. C. FIG. 25
shows transmission electron microscopy (TEM) image for GLY/AGE
nanoparticles with 7% crosslinking, after running reaction for 12
hours in methanol at 65.degree. C. FIG. 26 shows transmission
electron microscopy (TEM) image for GLY/AGE nanoparticles with 7%
crosslinking, after running reaction for 12 hours in methanol at
45.degree. C.
(4) Nanoparticle Formation Through Thiolene-Click Reactions Using
GLY/GEA Copolymers (50 nm)
[0315] To a round bottom flask equipped with a stir bar, was added
the GLY/GEA polymer (108.5 mg; 6.09.times.10.sup.-2 mmol; 1 eq),
3,6-dioxa-1,8-octanedithiol (11.11 mg; 6.09.times.10.sup.-2 mmol; 1
eq), and methanol (18.80 mL; 3.24.times.10.sup.-3M). The flask was
then fitted with a reflux condenser and lowered into an oil bath at
45.degree. C. to reflux for 12 h. The resulting solution was then
transferred to 10K dialysis tubing and dialyzed for 72 h against
methanol to remove any unreacted starting material. The remaining
product solution was then concentrated into a preweighed vial and
stored in the fridge. Yield: 28.9 mg (23.92%). .sup.1H-NMR (600
MHz, CDCl.sub.3) 6: The significant change is the reduction of the
allyl peaks at 5.72 and 5.04 ppm and the appearance of signals at
3.65 and 2.69 ppm corresponding to the protons neighboring the
thiols of the PEG linker after cross-linking All other aspects of
the spectrum remain similar to the polymer spectrum.
(5) Nanoparticle Formation Through Amine-Epoxide Reactions Using
VL/AVL/EVL Copolymers
[0316] To a round bottom flask equipped with a stir bar, was added
the VL/AVL/EVL polymer (30.1 mg; 2.45.times.10.sup.-2 mmol; 2 eq),
2,2'-(Ethylenedioxy)bis-(ethylamine) (2.717 mg;
1.83.times.10.sup.-2 mmol; 1.5 eq), and dichloromethane (7.54 mL;
3.24.times.10.sup.-3M). The flask was then fitted with a reflux
condenser and lowered into an oil bath at 45.degree. C. to reflux
for 12 h. The resulting solution was then transferred to 10K
dialysis tubing and dialyzed for 72 h against dichloromethane to
remove any unreacted starting material. The remaining product
solution was then concentrated into a preweighed vial and stored in
the fridge. Yield: 15.5 mg (47.4%). .sup.1H-NMR (600 MHz,
CDCl.sub.3) .delta.: The significant change is the reduction of the
allyl peaks at 2.96, 2.75, and 2.47 ppm and the appearance of
signals at 3.5 and 2.89 ppm corresponding to the protons
neighboring the secondary amine of the PEG linker after
cross-linking All other aspects of the spectrum remain similar to
the polymer spectrum.
(6) Nanoparticle Formation Through Amine-Epoxide Reactions Using
VL/AVL/EVL/OPD Copolymers
[0317] To a round bottom flask equipped with a stir bar, was added
the VL/AVL/EVL/OPD polymer (83.6 mg; 5.67.times.10.sup.-2 mmol; 2
eq), 2,2'-(Ethylenedioxy)bis-(ethylamine) (6.30 mg;
4.25.times.10.sup.-2 mmol; 1.5 eq), and dichloromethane (17.49 mL;
3.24.times.10.sup.-3M). The flask was then fitted with a reflux
condenser and lowered into an oil bath at 45.degree. C. to reflux
for 12 h. The resulting solution was then transferred to 10K
dialysis tubing and dialyzed for 72 h against dichloromethane to
remove any unreacted starting material. The remaining product
solution was then concentrated into a preweighed vial and stored in
the fridge. Yield: 72.6 mg (79.06%). .sup.1H-NMR (600 MHz,
CDCl.sub.3) .delta.: The significant change is the reduction of the
allyl peaks at 2.96, 2.75, and 2.47 ppm and the appearance of
signals at 3.5 and 2.89 ppm corresponding to the protons
neighboring the secondary amine of the PEG linker after
cross-linking All other aspects of the spectrum remain similar to
the polymer spectrum.
7. Preparation of Combination of the Dual Two Component Drug
Delivery System
a. General Procedure (for 17 Injections (20 .mu.L Per
Injection))
[0318] Bone morphogenetic protein (BMP2, 17 .mu.L, 10 mg/mL in 20
mM acetic acid) was added to 159 mg polyglycidol. Nanoparticles
containing MEK inhibitor (6.4% wt/wt) were dissolved in 244 .mu.L
sterile PBS to make 0.236 mg/mL solution with respect to MEK
inhibitor. Nanoparticle-MEK (4% crosslinked nanoparticle and 13%
loading of inhibitor) inhibitor solution was added to the
polyglycidol and BMP2 mixture and sonicated to yield a viscous, but
injectable solution (final polyglycidol concentration is 0.466
g/mL, each injection contains 10 .mu.g BMP2 and 3.4 .mu.g MEK
inhibitor).
8. Preparation of Reconfigurable and Responsive Network Systems
(1) Non-Functionalized Polyglycidol-Based Crosslinking Materials
for Hydrogels: as Fillers in Hydrogels and as Component in
Reconfigurable and Responsive Network Systems
##STR00175##
[0320] A mixture of poly(MEC, MAC) (100 mg, M.sub.n=4,700 g/mol,
0.10 mmol alkene), polyglycidol (100 mg, M.sub.n=6,000 g/mol), and
2,2-dimethoxy-2-phenylacetophenone (DMPA, 5.4 mg, 0.02 mmol) was
dissolved in DMF (0.10 mL) and allowed to stir at room temperature.
3,6-dioxa-1,8-octane-dithiol (17 .mu.L, 0.10 mmol) was added and
reaction was exposed to UV light (365 nm) for 5 minutes. The
resulting gel was washed in sequence with water, methanol, and
dichloromethane and allowed to dry overnight in vacuo to yield a
slightly opaque gel.
(2) Preparation of Polycarbonate/Polyglycidol Hydrogel Formation
Via Thiolene Click and Zinc Acetate Rearrangement
##STR00176## ##STR00177##
[0322] A mixture of poly(MEC, MAC) (100 mg, Mn=4,700 g/mol, 0.10
mmol alkene), polyglycidol (100 mg, Mn=6,000 g/mol), and
2,2-dimethoxy-2-phenylacetophenone (DMPA, 5.4 mg, 0.02 mmol) was
dissolved in DMF (0.10 mL) and allowed to stir at room temperature.
3,6-dioxa-1,8-octane-dithiol (17 .mu.L, 0.10 mmol) was added via
microsyringe, followed by the addition of zinc acetate (5.8 mg,
0.03 mmol). The reaction was exposed to UV light (365 nm) for 5
minutes. The resulting gel was then placed in a 120.degree. C. oil
bath overnight. The product washed in sequence with water,
methanol, and dichloromethane and allowed to dry overnight in vacuo
to yield a light yellow gel.
9. Preparation of Functionalized Polyglycidols
a. General Procedure
[0323] All reagents and solvents were commercial grade and purified
prior to use when necessary. Tetrahydrofuran was dried by passage
through a column of activated alumina as described by Grubbs
(Pangborn, A. B Organometallics 1996, 15, 1518-1520).
Dimethylformamide was distilled over CaH.sub.2 and stored over
molecular sieves. Glycidol was distilled under vacuum and stored
over molecular sieves. Thin layer chromatography (TLC) was
performed using glass-backed silica gel (250 .mu.m) plates and
flash chromatography utilized 230-400 mesh silica gel from Sorbent
Technologies. Size exclusion chromatography was utilized Sephadex
LH-20 from Sigma Aldrich. UV light, and/or the use of CAM and
potassium permanganate solutions were used to visualize
products.
[0324] Nuclear magnetic resonance spectra (NMR) were acquired on a
Bruker DRX-500 (500 MHz), Bruker AV-400 (400 MHz) or Bruker AV
11-600 (600 MHz) instrument. Chemical shifts are measured relative
to residual solvent peaks as an internal standard set to .delta.
7.26 and .delta. 77.0 (CDCl.sub.3), .delta. 3.31 and .delta. 49.0
(CD.sub.3OD). IR spectra were recorded on a Thermo Nicolet IR100
spectrophotometer and are reported in wavenumbers (cm.sup.-1).
Compounds were analyzed as neat films on a NaCl plate
(transmission).
(1) Preparation of N-Oxyphthalimide Polyglycidol Derivative
##STR00178##
[0326] Polyglycidol was synthesized according to known literature
procedure (Spears, B. R. Chem. Commun. 2013, 49, 2394-2396). To a
50 mL round bottom flask fitted with an argon balloon and
containing a solution of polyglycidol=2-3 kDa, 2.0 g) in DMF (25
mL) was added N-Hydroxyphthalimide (2.3 g, 14 mmol) followed by
triphenylphosphine (3.7 g, 14 mmol) at rt.
Diisopropylazodicarboxylate (2.7 mL, 14 mmol) was then added
dropwise and the resulting mixture was stirred at rt for 24 hrs.
The reaction was concentrated under reduced pressure and
precipitated twice in ether:ethyl acetate (1:1) to obtain 2.6 g of
the desired polymer as an off-white solid. IR (film) 3455, 3061,
2919, 1789, 1730, 1373, 1127, 731 cm.sup.-1; .sup.1H NMR (600 MHz,
CDCl.sub.3) .delta. 7.74-7.48 (br m, 4H), 3.06-4.49 (br m, 6H);
.sup.13C NMR (600 MHz, CDCl.sub.3) ppm 162.8, 134.7, 128.0, 123.4,
79.2, 77.8, 76.6, 74.9, 72.1, 71.4, 71.5-68.0 (br overlapping),
67.3, 65.2, 63.1, 61.2.
(2) Preparation of Aminooxy Polyglycidol Derivative
##STR00179##
[0328] To a 50 mL round bottom flask equipped with a stir bar and
an argon balloon was added a solution of N-oxyphthalimide
polyglycidol (1.0 g) in 1:1 mixture of methanol and dichloromethane
(25 mL). A 10 fold excess (based on measurements from the synthesis
of N-oxyphthalimide polyglycidol) of anhydrous hydrazine (4.5 mL,
140 mmol) was added and the reaction was allowed to stir for 12 hrs
at rt. The reaction mixture was filtered through 0.2 .mu.m PTFE
filter to remove the white solid byproduct. Further purification by
precipitation in ether followed by size exclusion chromatography
(sephadex LH-20 in methanol) yielded 800 mg of the desired polymer.
IR (film) 3407, 2873, 1373, 1113 cm.sup.-1; .sup.1H NMR (600 MHz,
CD.sub.3OD) .delta. 3.50-4.49 (br m, 6H); .sup.13C NMR (600 MHz,
CD.sub.3OD) ppm 79.9, 79.4, 78.6, 77.7, 74.7, 72.6, 72.0, 71.9-70.4
(br overlapping peaks), 70.0-68.8 (br, overlapping peaks), 66.2,
63.0, 61.9, 61.3.
[0329] As would be appreciated by those of skill, the percentage of
the amino-oxy can be readily adjusted and has been done for other
examples.
(3) Azide-Functionalized Polyglycidols and Allyl-Functionalized
Polyglycidols
(a) Preparation of Alkyne Functionalization of Polyglycidol
Method 1
[0330] A mixture of the appropriate propargyl bromide (1.00 equiv),
polyglycidol secondary hydroxyl group (1.00 equiv), dried potassium
carbonate (1.25 equiv), and 18-crown-6 (0.2 equiv) in DMF was
heated at 60.degree. C. and stirred vigorously under nitrogen for
24 h. The mixture was allowed to cool and add 50 mL methanol and
then remove the solid compound by vacuum filtration. The residue
crude product was precipitated in vigorously stirred acetone, which
was then decanted to afford the pure viscous product.
(b) Preparation of Alkyne Functionalization of Polyglycidol
Method 2
[0331] A dry flask was charged with propargyl bromide (0.36 equiv),
polyglycidol hydroxyl group (1.00 equiv), dried potassium hydroxide
pellets (1.08 equiv) in DMSO was stirred vigorously under nitrogen
at room temperature for 12 h. The mixture was diluted with 50 mL
methanol and the solid compound was removed by filtration. The
residue crude product was precipitated in vigorously stirred
acetone, which was then decanted to afford the pure viscous
product. As would be appreciated by those of skill, the percentage
of the amount of the azide group can be readily adjusted.
(4) Preparation of Glycidol-Alkyne-Azide
(a) Alkyne-Azide Click Reaction Catalyzed by Copper Foil
[0332] Polyglycidol-alkyne (0.11 g) was dissolved in 1 mL DMSO in a
microwave vial. Benzyl azide (0.061 mL) and Cu foil (0.25 g) was
added into the vial followed by irradiation at 160.degree. C. for
15 min. After completion of reaction, the reaction mixture was
twice precipitated into acetone and subsequently dried for 12 hours
under vacuum. The product was obtained as a highly viscous brown
liquid. FIG. 30 shows the Click reaction via NMR.
(5) Preparation of Random Copolyesters of .DELTA.-Valerolactone and
2-Oxepane-1,5-Dione
##STR00180##
[0334] To a 10 mL round bottom flask equipped with a stir bar and
an argon balloon was added isoamyl alcohol (37 .mu.L, 300 .mu.mol)
and tin(II) trifluoromethanesulfonate (1.3 mg, 3 mmol). The mixture
was stirred for 10 min. In a vial, flamed dried under vacuum, was
added 2-oxepane-1,5-dione (242 mg, 1.89 mmol, synthesized according
to literature procedure (Van der Ende, A. E. J. Am. Chem. Soc.
2008, 130, 8706-13)), .delta.-valerolactone (702 .mu.L, 7.57 mmol)
and 2 mL of N,N-dimethylformamide. Once all the 2-oxepane-1,5-dione
had dissolved, the solution was added to the reaction in one
portion, and stirred at rt for 24 hrs. The reaction was then
quenched with methanol and precipitated from hexanes to give the
desired golden brown polymer (806 mg, 80%). M.sub.w=1499 Da.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 4.42-4.24 (br m,
--C(O)CH.sub.2CH.sub.2O--), 4.14-3.95 (br m,
--CH.sub.2CH.sub.2CH.sub.2O--), 2.82-2.67 (br m,
--CH.sub.2C(O)CH.sub.2--), 2.63-2.46 (br m,
--OC(O)CH.sub.2CH.sub.2C(O)--), 2.38-2.18 (br m,
--OC(O)CH.sub.2CH.sub.2CH.sub.2--), 1.91-1.79 (br m,
(CH.sub.3).sub.2CH--), 1.72-1.52 (br m,
--C(O)CH.sub.2CH.sub.2CH.sub.2CH.sub.2O--), 1.52-1.42 (br m,
(CH.sub.3).sub.2CHCH.sub.2--), 0.91-0.82 (br m,
(CH.sub.3).sub.2CH--).
(6) Preparation of 4-Pentenoyl Polyglycidol
##STR00181##
[0336] To a flame dried 25 mL round bottom flask equipped with a
stir bar and argon balloon was added polyglycidol (1 g) and
pyridine (2 mL, 25 mmol). The reaction mixture was stirred at rt
for 10 min then cooled to 0.degree. C. Pentenoyl chloride (607
.mu.L, 5.50 mmol) was added dropwise to the reaction. The reaction
was allowed to warm up to rt and stirred for 12 hrs. Reaction was
then diluted with N,N-dimethylformamide (2 mL) and precipitated in
a mixture of diethyl ether and ethyl acetate (1:1) to give the
desired product as a pale yellow oil (520 mg, 52%). .sup.1H NMR
(400 MHz, CD.sub.3OD) .delta. 6.07-5.70 (br m, CH.sub.2.dbd.CH--),
5.21-5.01 (br m, CH.sub.2.dbd.CH--), 4.96-4.50 (br s, --OH),
4.40-4.03 (br m, --CHCH.sub.2OC(O)CH.sub.2--), 4.02-3.86 (br m,
--CHCH.sub.2OH), 3.82-3.22 (br m, --OCHCH.sub.2CHO--), 2.60-2.23
(br m, --C(O)CH.sub.2CH.sub.2CH.dbd.CH.sub.2), 1.71-1.66 (br m,
(CH.sub.3).sub.2CH--), 1.50-1.39 (br m,
(CH.sub.3).sub.2CHCH.sub.2--), 0.95-0.81 (br m,
(CH.sub.3).sub.2CH--). As would be appreciated by those of skill,
the amount and type of allyl group can be readily adjusted.
(7) Preparation of 3-Mercaptopropanoyl Polyglycidol
##STR00182##
[0338] To a 25 mL round bottom flask, flame dried and equipped with
a stir bar and argon balloon, was added a solution of polyglycidol
(1 g) in N,N-dimethylformamide (1 mL), 3-mercaptopropionic acid
(1.4 mL, 16.6 mmol), and p-Toluenesulfonic acid (34 mg, 0.2 mmol).
The mixture was stirred at 100.degree. C. for 24 hrs. The mixture
was diluted with N,N-dimethylformamide (2 mL), and precipitated in
ether. The product was further purified by size exclusion
chromatography (sephadex LH-20 in methanol) to yield the desired
polymer (200 mg). .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 4.93
(br s, --OH), 4.46-4.08 (br m, --CHCH.sub.2OC(O)CH.sub.2--),
4.01-3.81 (br m, --CHCH.sub.2OH), 3.85-3.41 (br m,
--OCHCH.sub.2CHO--), 2.81-2.63 (br m, --C(O)CH.sub.2CH.sub.2SH),
1.78-1.66 (br m, (CH.sub.3).sub.2CH--), 1.52-1.42 (br m,
(CH.sub.3).sub.2CHCH.sub.2--), 0.98-0.87 (br m,
(CH.sub.3).sub.2CH--). As would be appreciated by those of skill,
the amount and type of mercapto group can be readily adjusted.
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